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This is to certify that the

thesis entitled

USING STR ANALYSIS TO DETECT
PRIMARY DNA TRANSFER

presented by
Tara Ann Reinholz

has been accepted towards fulfillment
of the requirements for

\MLdegree in W03
with Specialization in
Forensic Science

 

Date b l

0-7539 M50 is an Affirmative Action/Equal Opportunuy Institution

 

 

 

Ll 63AM
Mich i9 ear; State

U n iversity l

 

 

 

E11 I BOX to remove this checkou I: from _y
PLACE m R AVRng FINES return on or before
TO

M

our l‘ECOrd.
Clare due

i f"(i‘QIJesterL

 

 

 

DATE DUE
DATE DUE

VV‘

 

DATE DUE

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

6/01 cJCIRClDateDue.p65.p_15

“x

USING STR ANALYSIS TO DETECT
PRIMARY DNA TRANsFER

BY

Tara Ann Reinhol z

A THESIS

Submitted to
Michigan State University

in partial fulfillment of the rec“l
for the degree of

irEmente
MASTER OF SCIENCE

School of Criminal Justice

2002

ABSTRACT

USING STR ANALYSIS TO DETECT
PRIMARY DNA TRANSFER

BY

Tara Ann Reinholz

In this study, the primary transfer of DNA from a
person’s hand to an object was explored. A hammer with an
unfinished wood handle was used as a test object, and
holding times were varied l) to determine if a relationship
existed between holding time and the amount of DNA
recovered and 2) to evaluate the sensitivity of the testing
nethod. An AmpFlSTR®>Profiler Plus Amplification Kit and
the ABI Prism®>310 Genetic Analyzer, equipped with
Genescan®>2.0.2 and Genotyper®>2.0, were used to generate
DNA profiles. In addition, two new procedures were
developed in an attempt to increase the sensitivity of the
testing method. The results Show that primary transfer of
DNA to this type of surface is common. It is possible to
generate complete DNA profiles from holding times as low as
5 seconds, and no apparent connection exists between
holding time and the amount of DNA recovered. The new
Procedures proved useful in increasing allele detection
but, occasionally, heterozygote peak imbalances of true

alleles were observed.

 

ACKNOWLEDGMENTS

First, I would like to thank my professor and advisor
Dr. Jay A. Siegel. His knowledge, guidance, and commitment
to forensic science have Shaped me into the scientist that
I am today. This project would not have been possible if
not for his extraordinary patience and support.

Second, I would like to thank the entire Biology Unit
at the Michigan Department of State Police Forensic
Laboratory in Northville, Mi. Lynne Helton, Heather
Spillane, Leslie Nixon, Dorothy Martus and Guy Nutter all
played an instrumental part in my research from the very
beginning. Without them, I would not have gained the
knowledge or had the resources needed in order to carry out
my project. I want to offer a special thanks to Heather
Spillane. Heather’s suggestions and ideas created the
backbone of my project, and her time, knowledge, experience
and incredible patience allowed me to successfully carry it
out. Without her, and the rest of the unit, none of this
would have been possible.

Third, I need to thank all of the individuals who were

a part Of my sample population. Without the donation of

their time, the Significance of the results obtained in

this study would have been severely compromised.

iii

TABLE OF CONTENTS

 

 

 

 

 

 

 

 

 

 

List of Tables v
List of Figures vi
List of Abbreviations vii
Introduction 1
Review of the Literature ' 9
Materials and Methods 12
Results and Discussion 39
Conclusions 64
Suggestions for Further Research 65
References 66
Appendix A - Electropherograms generated from
decontamination testing 72
Appendix B — Electropherograms generated from
known DNA samples 76
Appendix C — Electropherograms generated from
5, 15, and 30 second samples 90
Appendix D — Electropherograms generated from
samples containing 3ul of
PCR Product 136
Appendix E - Electropherograms generated from
samples containing 3ul of
concentrated PCR product- 152
Appendix F - Electropherograms generated from
168

 

 

 

 

 

ladders and quality control samples .........

iv

LIST OF TABLES

DNA
. marl
Table 1 - Applied BlOSYStemS Quant iBlOt® H”
QuantitatiOn Klt Contents.
Profiler plus PCR
Table 2 —— Applied Biosystems AmpFlSTR®
Amplification Kit: contents.
. iler Plus
Table 3 — Applied Biosystems AmpFlSTR® PIGi
loci.
Table 4 - 310 run parameters.
Table 5 — Stutter percentages.
Table 6 _. First round quantiblot IesLths
__ Second round quantiblot
Table 8 — DNA profiles Of known Samples
Table 9 — DNA profiles of 5, 15, and 30
&%Q 1e8-
Table 10 — Profile types of 5, 15, and 30 0nd Sam?
Table 11 — DNA profiles of samples USing 3 econd samples.
PCR product. K11 of
Table 12 — DNA profiles Of Samples using 3
of concentrated PCR product, Lll
Table 13 - Profile types of samples using

3111
PCR product and 3ul of concent:bat8dc>;C
product. R

 

Figure 1

Figure 2 -
Figure 3 -

Figure 4 ~

Figure 5 —
Figure 6 —

Figure 7 —

 

LIST OF FIGURES

DNA struc : ure.

Polymerasez chairl reaction-
Capillary electrophoresis-
Yield gel interpretation-

Quantiblot interpretat i on -

- ers-
Genescan® analysst paramep

Laboratory deSign.

vi

 

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MIA
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A-S '-
A-lS -

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CE _
DNA __
FBI -
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LCN ——
MSP —
ng -
PCR —-

QA _

QB _
QC _

RFU
STR
SWGDAM -

TAE
TE
TWGDAM _
ul -

 

LIST OF ABBREVIATIONS

l
ndi'v'iailual

. . n i
letter deSlgnation “A" glven to a

who took part in the study

individual “A” / 5 second hOld

individual. “A” / 15 second hold
individual. “A” / 30 secol’ld hold tor Dire
American Society of Crirne Labora
capillary electrOphores i5
dOXyribonucleiC acid

Federal Bureau of Investigatiofl
iI'Iternal laboratory contrOl

low copy nurnber . 08
Michigan Department of State POl )—
nanOgram

Polymerase Chain react ion
picogram

quality assurance

gnantiblot

Quality ContrOl

relative fluorescent Lll‘iit

short tandem repeat

Scientific working grOL1p on DNA anal
methods YSI

Q
trig/acetate/EDTA
tris/EDTA

Technical working group on DNA '31")
31y .
81

microliter S meth
(Dds

vii

 

 

 

 

11'

 

I NTRODUCT I ON

DNA (Deoxyribonucleic acid) represents one

makeup. Located in the nucleus of cells!

for how the human body funCtions, as Well as

DNA it i5
Forensicalxy

tec‘fiflo

this 0 ' 5% di f ferenCe that is import: ant _ TOéaY' ‘ 3x30

«“3

appearance . Approximate 1y 9 9 . 5 % o f the

identical 111 all human rpeifnas [131

makes it possible to detect these di

0 . . . .
ne indiVidual to be d1. fferentiated from a .96
. (,6 ,0 6
Common sources of nuclear DNA lnClude Why 0 A36
cells .300 Cf,
r spermatona, tissues and hair rootS. 9 O
9'
biological con - eé OI
stituents are commonly encount: €335 COOXi
D16

scenes, DNA has proven to be an eEritrernely va lag

the criminal justice system.

Source. _ . .
A hammer Wlth an unfinisth wood ha d1 5’ DNA
b a ' - . , e w
Y n individual for a SpeCific amQunt of tin Q
sample Wa e’ a!) field
S Collected from the handle Th'
t f . lS tYpe a
ran
S Er, DNA from ones hand to an Object i of
I S
as a ‘ I‘
primary transfer. Before the next ind . eferred
l jdu t
a1 held

Onto the
handle, it was decontaminated in O
eliminate a bGEr
my DNA left over from the previQ to
Lls
hOlder.

Choosin
9 t0 decontaminate before each hold

$11-
m e
J‘I‘lated the

 

 

 

 

A]?

161 ant.
need to use separate hammers for each part p 0
W'
for C
- - hosen
unfinished wood surface Was speCificallY c on
m
com

- many
reasons. One, this substrate 18 found Or1

on5
household items that could be used as weap

d
knives, baseball bats, etc. Two, un’fln oecu

. mote
rougher surface Which could p0881bly pro

. {a 8
of DNA transfer. A DNA profile was gene P» 019

fl
. . , know
sample and compared to the indiVidualS

. 0V8 "O o
the two profiles matched: this would Pr

transfer of DNA to this type of subst

. 0 8
This reSEEarch has the potentlal to p

. 6 a“
a Knj’fi 0‘9
area of forensic evidence. For example:
. . w“ dany
crime scene exhibiting only a partial finger? 559

roc
could not be identified, WOUld nOt usually be P

further_ BY incorporating the procedures pr Cposed Jill HHS

study, the knife could become a very probati—Ve pieQ

. G
evidence. The appearance Of the partial prL rut Pro Of

an individual did, in fact, touch the handle of U3 88 “Pat
Q

BY C01lecting a sample from the handle, a DNA PrOf knife

1'16 '

be generated and a possible SUSPeCt could b3 , co
lden , “Id

In addition to opening a new area of fgr ied.
Ens.
. 1c
ev1dence, this study explores the need to acid
res
. . . s .
of contami nation The POSSlblllty Of detth. the lSsue
lng Dr

imary DNA

transfers reinforces the importance of weabi
r19
gloves

7.

 

 

 

 

T176

during the collection and handling of all eVldenCe-

tefltial

- . - ' the PO
forenSlC seience community needs to reallze ‘tivitY
eflsl

for contamination to Occur. Wlth the lncre cies
t agen
an

of the current testing methods, law en «161 o
e

. t
and laboratories need to fOllow the higheg oi the
in order to insure the reliability a

testing performed.

PolymeraseW ‘ €569 6”

_ C
DNA 18 a double st: randed mo]_EBC2Llle tha

6
itfi
nucleus of the cell. It: is composed of 4 d 6

Adenine (A) , Thymine (T) , Guanine (G) I

It is the specific order of these bases along 66
ps3

strand that dEtermineS one’S genetic makeup-

one

Figure 1’ two of these bases Specifically at: Each to

another to fOrm a base pair [19] ~ A binds with T

binds with C, It is estimated that the humaI—j

genome Consists of over three bill ion of the Se 13
‘38

[15] . pairs

AS mentioned earlier: apprOXimatelY 0~ 3 9..

Q

of
' . t
bllllon baSe pairs differ from one person to he 3

t:
be next_
and
Polymorphi Sm is a short: tandem repeat, or ST. One tYpe
R

These differences are called p01ym0rphisms,

0f

7 . ‘ 8
base pair repeats thalt Occur along a DNA TRS are 2‘

trand [4] .

 

 

 

Structure of DNA

  
    
 

backmniese

Base

   

.hala ”#6‘
?T\\__._~\N‘_
Hydrog e :n
ban
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Figure
1 _
DNA structure-

M

' e the
Forensically, the tetranuCleotlde repeats ar

current areas of intereSt. For example. core

‘5
or locus, on a DNA strand, an STR c20111d exl exist
alleles, to

repeat unit of TAGC. Different forms. or

amongst the population, thus allowing f0
be differentiated from another. One p8 ext
'0
30 allele (TAGC repeated 30 times)
40 allele (TAGC repeated 40 times) ° . m.
rue
longer than the other, this type of
referred to as a length polymorphism- 096

9
as two pu‘ 6&6

The POlYmerase chain reaction h (0

forensic DNA analysis. One, is to ampllfy' 01;

millions of. copies of the particular STR Of
doing this, DNA that has become decomposed due
to environmental insults has the pOssibilitY
viable sample. Also, very small Samples can be €115l
Two, is to attach a fluorescent label to the STR fQ .
detection by a laser- By using Primers iii-199% d with ‘5
different Colored labels, many STR locations

DNA strand can be amplified at one time, thus ’

much higher power of discrimination. FigurQ

the process of pCR [22]

Ca illar Electro hores iSQCE)

of Dec:

M

Polymerase Chain Reaction

‘.

firegion 0f irzeresz. D131 .‘5 dena tured. 9r;.-.:r MLLL
toeach Strand. Pr Lnev. B213: strand
is syn hes; zed uehm n31 primers an
e1 :h ....r: 2:: srari.

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denatured, {gr-mfg are denature" , primrs are d: natured, p." .1: 3:5
atzached, and he attached: and :he G333 iE-d and the.
nut-her of [3:92, nutter oi Eli-‘4. number of Jr’s-"A \’
Strands are doubled. strands are d‘mmd' 3-8170, 5555‘ d3 11b ..

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r M? ' . " "3315' -'
Lagma- 301211325 the Emmi-CE “i r S

 

Figure 2 \ Polymerase chain reaction.

6

 

H)

Electrophoresis is defined as a technique used for the
separation of molecules by means of electric current [1].
Molecules that have similar charges but differ in size can
be separated using this technique. Since DNA is
negatively charged and the STRs vary in length from one
person to another, this makes capillary electrophoresis
a perfect technique to use for STR analysis.

As diagrammed in Figure 3, a sample is pulled up into
the capillary and the applied electric current allows the
negatively charged DNA fragments to travel from the
negatively charged cathode to the positively charged anode
[6]. As the fragments travel through the capillary they
are separated by size. The smaller sized STRs reach the
anode first, followed by the larger fragments. When
referring to the example given earlier, the 30 allele would
reach the anode in front of the 40 allele because it is a
smaller fragment. As they pass through a detector window,
a laser excites the fluorescent tag that was attached to he
STR during PCR, and a CCD camera collects the emitted
fluorescence. Computer programs analyze the data
and a genetic profile is generated for that particular

sample.

 

 

 

 

Com uteI
P in»

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Buffer

 

 

 

 

 

 

R ,
Cathode I.“ i-xIlOdE-B
“1} +

 

 

 

Time {min}

 

 

Figure 3 — Capillary electrophoresis.

REVIEW OF THE LITERATURE

In 1997, Roland A.H. Oorschot and Maxwell K. Jones
were the first to explore the possibility of using PCR/STR
technology to detect primary and secondary DNA transfers
[24]. Although the holding times were lengthy (up to 20
minutes), it was proven that the possibility to transfer
DNA from ones hand to an object (primary transfer), as well
as from an object to ones hand (secondary transfer) does
exist. The substrates tested included leather, plastic,
and glass. The human hand itself also proved to be a
successful substrate. After a one minute handshake, the
palm of one participant revealed both DNA profiles.
Oorschot and Jones’s study set the stage for all future
research in this area.

Colleagues at the Connecticut State Police Forensic
Laboratory duplicated the Oorschot/Jones study, but
concentrated more on secondary transfers [16]. Skin to
skin to object and skin to object to skin were the two
modes of secondary transfer that were explored. In
contrast to Oorschot and Jones study, the handling times
were greatly decreased, thus resulting in significantly
lower yields of DNA. The results of their study indicate

that primary DNA transfer is possible but not always

detectable, and, unlike Oorschot and Jones study, secondary
transfers were not able to be detected.

In research conducted by R.A. Wickenheiser of the
Royal Canadian Mounted Police (RCMP), PCR/STR technology
detected the transfer of DNA to over 15 different
substrates [28]. Because of these findings, this approach
was implemented into everyday casework, and a recent murder
case was solved. The handle of a knife recovered at a
crime scene revealed the DNA profile of the suspect [27].

P. Wiegand and M. Kleiber of the Martin-Luther—
Universitat in Germany used PCR/STR technology to detect
the transfer of DNA from the hands of the suspect onto the
neck of the victim in cases of strangulation [29]. 16
suspect/victim combinations were used and the success rate
was over 70% for the three STR loci tested.

Colleagues at the University of Gent in Belgium
performed DNA profiling on physical fingerprints left on
glass and wooden plates [23]. Good results were obtained
from both substrates when at least 5 physical fingerprints
were present.

At the 1999 proceedings of the National Commission on
the Future of DNA evidence, Lynne Fereday of the Forensic
Science Service in England offered a summary of the past

research in this area [7]. In this summary, many of the

10

studies conducted by the authors mentioned above were
discussed. In addition, the Forensic Science Service is
currently conducting their own DNA transfer studies. These
studies were briefly described, but have yet to be
published. One experiment involved determining the most
recent driver of a vehicle by detecting DNA transferred to

the steering wheel by the driver’s hands.

11

MATERIALS AND METHODS

Decontamination Testing

 

Before an individual was to touch the handle of the
hammer, it was decontaminated in order to insure that
DNA had not been left over from a previous holder. The
handle was soaked in a bleach/distilled water solution
for 20 minutes, wiped off with a paper towel, rinsed in
distilled water, wiped off with a paper towel, placed in a
heated oven to dry, and allowed to come to room
temperature. Both a 10% and 20% bleach/distilled water
solution were tested and the 20% solution proved to be the
most effective.

This entire procedure was extensively tested in order
to insure that no DNA carry over was taking place. A male,
who consistently transferred DNA, held onto the handle of
the hammer for 2 minutes. The handle was decontaminated,
followed by a 2 minute hold by a female. Procedures, that
will be explained in more detail shortly, were followed in
order to generated DNA profiles from the female samples.
The absence of a male/female mixture in the female samples
will assure that the decontamination process is working

properly.

12

First Round of Sample Collection

 

Initially, samples were collected from 32 Caucasian
males and females, 16 years and older. In order to give a
true representation of primary DNA transfer occurring in
this type of population, the hands of the individuals were
not controlled in any way. Not controlling the hands
refers to not requiring any special procedures, such as
washing the hands or rubbing them together, before coming
into contact with the handle. By encouraging normal,
everyday behavior by the test subject, evaluating the rate
at which primary transfer occurs can be more accurately
determined.

After decontamination, an individual held onto the
handle of the hammer for 2 minutes. Forcefully gripping
onto the handle, swinging the hammer around, and switching
hands were all encouraged, thus mimicking actions that
could occur during the commission of a crime with this type
of weapon. After 2 minutes, a sample was collected from
the handle by using a technique known as the double swab
technique [20]. First, a sterile cotton tipped swab was
dipped into distilled H20 and rubbed over the entire handle
of the hammer. Second, a dry swab was used to do the same.
Both swabs were allowed to air dry before placed in a

storage tube. This storage tube was given a letter

13

designation in order to identify the individual who

contributed the sample. All samples were stored in a -20°C

freezer until time to be extracted.

Organic Extraction

 

For each sample, a scalpel was used to remove the
cotton tip from each swab and both were placed in a single
extraction tube. Six—hundred microliters of stain
extraction buffer (10 mM Tris-HCL, 10 mM EDTA, 50 mM NaCl,
2% SDS, pH 7.5) was added and the tube was placed in a
centrifuge and spun down (5 minutes/15,000 9). Thirty
microliters of Protinase K (10 mg/ml) were then added. The

tube was vortexed to mix, spun down(5 minutes/15,000 g) and

placed in a 56°C oven for an overnight incubation.

The next day, the cotton tips were removed from the
extraction solution and placed in a basket insert. The
basket was inserted into the original extraction tube, and
and this was spun down (5 minutes/15,000 g). The basket
insert was removed from the extraction tube and thrown
away. Five—hundred microliters of a Phenol /Chloroform /
Isoamyl Alcohol solution (25:24zl) were added and the tube
was vortexed to mix for 5 - 10 seconds. After being spun
down (5 minutes /15,000 g), the DNA extract (top layer),

was removed and placed into a new microcentrifge tube.

14

Next, Amicon®>centricon concentrators were used to
purify the DNA extract. Five-hundred microliters of
Tris/EDTA (TE) Buffer (10 mM Tris-HCL, 0.1 mM EDTA, pH 8.0)
were added to the centricon filter and it was spun down
(500 g/10 minutes). The rinse filtrate in the centricon
reservoir was discarded. One milliliter of TE Buffer and
the entire DNA extract were added to the centricon filter
and spun down (500 g/30 minutes). This procedure was
performed three times. After the third wash, the centricon
filter was removed from the reservoir and inverted into a
vial. This was spun down (3 minutes/1000 g) and the
filtrate was removed from the vial and placed into a new

microcentrifuge tube. The purified DNA extract (~ 30 ul)

was stored at ~20°C until further testing.

Yield Gel

A yield gel can be used to determine the quality of
DNA and determine how much DNA is present in an extract. A
submarine gel electrophoresis unit was used to carry out
this procedure. The running buffer, 25 ul of ethidium
bromide (0.5 ug/ml) in 250 ml of Tris/acetate/EDTA (TAE)
buffer (40 mM Tris—acetate, 1 mM EDTA, pH 8.3), was poured
into the apparatus, and a 1% agarose gel (1.3 grams

agarose/130 ml TAE buffer), containing two columns of 14

15

wells, was placed into the buffer.

Wells # l - 7 in the first column were reserved for
standards to which the samples are compared. Three
microliters of Lambda Hind III/Eco R1, a mixture of
standards, were added to well #1 in columns 1 and 2, and 6
ul of the following DNA standards were added to wells #2-7
in column 1: 500 ng, 250 ng, 125 ng, 63 ng, 31 ng, and 15
ng. For each sample, 4 ul of the purified DNA extract were
added to 2 ul of 6x loading buffer (bromophenyl
blue/glycerol) and this 6 ul mixture was added to a well.
Electrophoresis was carried out for 12 minutes at 175
volts.

An ultraviolet (UV) light box was used to obtain a
photograph of the gel. The ethidium bromide that was added
to the running buffer binds to the DNA strand and
fluoresces in the presence of UV light. Therefore, the DNA
in the gel can be visualized when exposed to ultraviolet
light. The picture was then used to evaluate the DNA
sample.

When interpreting a yield gel (Figure 4), the samples
are compared to standards in order to determine the quality
and quantity of the DNA sample [2,17]. A solid band
represents high molecular weight, or good quality, DNA and

a smear represents degraded, or poor quality, DNA. Quantity

16

Solid, bright band = large quantity of high
molecular weight DNA

Solid, light band = small quantity of high
molecular weight DNA

 

Smear = degraded DNA

 

Figure 4 - Yield gel interpretation.

17

is determined by picking a standard that most closely
resembles the intensity of the sample band. The brightness
of the band is directly proportional to the quantity of
DNA. If a band is not seen, a more sensitive quantitation

method will need to be performed.

Applied Biosystems QuantiBloUO Human DNA Quantitation Kit

A QuantiBloUO kit was used as a more sensitive way to
quantitate the purified DNA extract (Table 1). Yield gels

detect from 125 ng of DNA/ul to 3.75 ng of DNA/ul, while

the QuantiBlotC>detects from 2 ng of DNA/ul to 0.03125 ng
of DNA/ul. This test is based on the hybridization of a
biotinylated, primate—specific probe (D17Z1 locus) to DNA
samples immobilized on a nylon membrane and
chemiluminescent detection of the bound probe. The
procedures for hybridization, chemiluminescent detection,
and interpretation of results printed in the product insert
were followed [5].

Just as in a yield gel, the samples were compared to
standards (10 ng/5 ul to 0.15 ng/S ul) to in order to
determine the concentration of DNA present in the samples.
When interpreting the quantiblot (Figure 5), the size and

intensity of the bands on the X—Ray film were evaluated

[3].

18

Table 1 - Applied Biosystems QuantiBloUO Human DNA
Quantitation Kit contents.

 

 

 

 

 

 

 

 

Reagent VOlume Description
1 vial containing 1pmole/ul
_ biotin lated DNA oli onucleotide
QuantiBlotC> 220 ul y “g
D17Z1 P b probe of the follow1ng sequence:
r0 e 5’—biotin-TAGAAGCATTCTCAGAAACTA
CTTTGTGATGATTGCATTC-B’.
1 bottle containing Horseradish
Enzyme . . .
. Perox1dase—Streptav1din (HRP-SA)
Conjugate: 2 0 ml . . .
conjugate supplied in buffer
HRP-SA . .
with preservative.
Bromothymol 1 vial containing 0.04%
. 200 ul .
Blue Solution Bromothymol Blue in water.
QuantiBlot®>DNA. 250 ul 1 vial containing 2 ng/ul human
Standard A genomic DNA in TE buffer.
QuantiBloU® DNA. 100 ul 1 vial containing 0.7 ng/ul
Calibrator 1 human genomic DNA in TE buffer.
QuantiBloU® DNA 100 ul 1.vial containing0.1 ng/ul human

Calibrator 2

 

 

genomic DNA in TE buffer.

 

19

 

10ng

2.5

1.2

0.3

0.15

Figure 5 - QuantiBlot® interpretation

Hill!

C» '“‘-

11 Ill

20

| Y I

10ng

()1

2.5

1.2

0.6

0.3

0.15

Second Round of Sample Collection

 

The results generated from the quantiblot procedure
allowed the samples to be put into groups according to the
amount of DNA present in the purified DNA extract. Because
of time and resource limitations, approximately half of the
individuals from each group were randomly chosen to take
part in the second round of sample collection. Three
samples, one at 30 seconds, 15 seconds and 5 seconds,
were collected from each person. A buccal sample (cells
from the inside of the mouth) was also collected from each
participating individual so a known DNA profile could be
generated. These samples were exposed to all of the same
procedures mentioned above. The letter designations
previously given to the individuals were used to identify
the buccal samples. The letter designation followed by a
dash and the handling time in seconds, was used to identify

the timed samples.

Applied Biosystems AmpFlSTR® Profiler PlusTM PCR
Amplification Kit

 

 

PCR was carried out using the Profiler Plusm‘Klt

(Table 2) and the Applied Biosystems GeneAmp®>2400 PCR
Instrument System [3]. This kit will co—amplify nine STR

loci and a segment of the amelogenin gene for gender

21

Table 2 — Applied Biosystems AmpFlSTRC>Profiler Plusm'PCR
Amplification Kit contents.

 

Kit Component

Volume

Description

 

AmpFlSTR® PCR
Reaction Mix

1.1 ml/tube

Two tubes each containing MgClL
deosynucleoside triphosphates

(dATP, dCTP, dGTP, dTTP), bovine
serum albumin (BSA), and 0.05%
sodium azide (NaN3) in buffer and

salt

 

One tube of locus—specific 5—FAM—
, JOE-, and NED-labeled and
unlabeled primers in buffer to

 

 

 

 

 

 

 

AEPFISTRC) amplify the STR loci D3Sl358,
Pr0f¥1er PlusTM 1‘1 ml vWA, FGA, 0881179, 021811,
Primer SEt 018851, 058818, 0138317, and
D78820, and the gender marker
amelogenin
AmpliTaq GoldC> Two tubes of enzyme with an
DNA Polymerase 50 Ul/CUbe activity Of 5 U/Ul
One tube containing 0.10 ng/ul
human cell line DNA in 0.05% NaN3
and buffer. The enot e of this
AmpFlSTRC) female DNA is D3SI358 15,15; vWA
contrOI DNA 0'3 ml 17,18; FGA 23,24; 0881179 13,13;
9947A 021811 30,30; 018851 15,19;
D58818 11,11; D13S317 11,11 and
D78820 10,11
Mineral oil 5 ml One dropper bottle
One tube of AmpFlSTRC>Blue
AmpFlSTRC>B1ue 25 1 Allelic Ladder containing the
Allelic Ladder u follOWing amplified 5-FAM-labeled
alleles: D3S1358 12-19, vWA 11-
21, FGA 18-30 (including 26.2)
One tube of AmpFlSTwO Green II
Allelic Ladder containing the
following amplified JOE-labeled
AmpFlSTw® Green alleles: amelogenin X and Y,
II Allelic 25 U1 D881179 8-19, D2181l 24.2-38
Ladder (including 28.2, 29.2, 30.2,
31.2, 32.2, 33.2, 34.2, 35.2),
D18851 9-26 (including 10.2,
13.2, 14.2)
One tube of AmpFlSTRC>Yellow
AmpFlSTR®> Allelic Ladder containing the
Yellow Allelic 25 ul following amplified NED-labeled
Ladder alleles: D58818 7—16, D138317 8—

 

 

15, and D78820 6-15

 

22

 

identification (Table 3). Prior to amplification, the DNA
samples were diluted to the appropriate concentration.
According to studies done by the Michigan Department of
State Police (MSP), approximately 1.0 ng of DNA is required

for optimum amplification to occur. The purified DNA

extract was diluted in Milli-Q®>water to a final
concentration of 0.1 ng/ul, and 10 ul of this DNA stock
solution would be added to the PCR reaction. If the DNA
extract did not contain a high enough concentration of DNA,
10 ul of straight extract was used.

Each PCR reaction required 10.5 ul of reaction
mixture, 5.5 ul of primers, and 0.5 ul of AmpliTan>Gold
DNA polymerase. After the number of samples to be
amplified was determined, a master mix of these reagents
was prepared in a separate microcentrifuge tube. Fifteen
microliters of this master mix was added to a PCR reaction
tube, followed by 10 ul of the 0.1 ng/ul DNA stock solution
prepared earlier. This gave a final reaction volume of 25
ul. PCR was then carried out using the following

parameters [3]:

1. Initial Denaturationzx3> 95°C/1l minutes
Denature = 94°C/1 minute
2. Step Cycles:>:>:>:>:>:>:> Anneal = 59°C/l minute

(28 cycles) Extend 72°C/1 minute

23

Table 3 — Applied Biosystems AmpFlSTR®>Profiler Plusm‘loci.

 

 

 

 

 

 

 

 

 

 

 

 

Locus Chromosome Common Sequence 128;: ea Dye
Designation Location Motif (88?“ Label
TCTA (TCTG) 1-3
- - F
D381358 3p (TCTA)n 114 142 5 AM
TCTA(TCTG)}4
_ 7 _ _
VWA 12p12 pter (TCTA)n 15 197 5 FAM
(TTTC) 3TTTTTTCT
FGA 4q28 (CTTTWnCTCC(TTC 219—267 S-FAM
C) 2
. X: p22.1-22.3 - 107
Amelogenin Y: p11.2 _ 113 JOE
D881179b 8 (TCTR);C 128—168 JOE
[321811 (TCTA)n(TCTG)n[(
21 TCTA)3TA(TCTA)3T 189-243 JOE
CA(TCTA)2TCCATA
] (TCTA) n
D18SSI 18q21.3 (AGAA)n 273-341 JOE
D58818 5q21—31 (AGAT)n 135—171 NED
D13S317 13q22-31 (GATA)n 206—234 NED
D78820 7q11.2l—22 (GATA)n 258-294 NED

 

 

 

 

 

a. The size range is the actual base pair size of sequenced

alleles contained in the AmpFlSTR®>Profiler PlusTM
Allelic Ladders. The sizes in the table include the 3’

A nucleotide addition.

b. In some literature references,
as D6SSO2

c. R can represent either an A or G nucleotide.

24

this locus is designated

 

3. Final Extension2>2>::>: 60°C/45 minutes

4 . Final Step:>:>:>:>:>:>:>:> 2 5°C/ forever

ABI PrismC>310 Genetic Analyzer

 

The 310 Genetic Analyzer was used to perform capillary
electrophoresis on the amplified samples. First, a
capillary electrophoresis reaction mixture was prepared.
This was composed of 24 ul of deionized formamide and 1 ul
of GeneScan—SOO [ROX] internal lane size standard for each
sample. Just as in PCR, the number of samples to be run on
the instrument was determined and a master mix was prepared
in a separate microcentrifuge tube. Twenty—five
microliters of master mix were added to a 310 sample tube,
followed by 1 ul of amplified product. The ladder sample
was prepared by adding 3 ul of.AmpFlSTRC>Profiler PlusTM
allelic ladder to 25 ul of the master mix. This ladder
consists of DNA fragments of known sizes to which the
unknown samples will be compared to. Samples were then
incubated at 96°C for three minutes and cooled in a benchtop
cooler for three minutes.

After appropriate electrophoresis parameters were
selected, the samples were loaded onto the instrument and

the run was started (Table 4). Following separation,

GenescanC>2.0.2 and GenotyperC>2.0 software were used to

25

Table 4

— 310 run parameters.

 

Injection Time

5 seconds

 

 

 

Injection kV. 15.0
Run kV. 15.0
Run Temperature 60°C

 

Run Time

24 minutes

 

Matrix File

Profiler plus

 

Size Standard

ROX GS 500

 

Instrument Configuration

Pop-4TM polymer with.1—nd
syringe

 

 

Instrument Module

GS STR POP4 (1ml) F

 

 

26

 

analyze the raw data and electropherograms, or genetic

profiles, were generated. Figure 6 illustrates Applied

Biosystems recommended GenescanC>analysis parameters [3].
For this study, the peak amplitude thresholds illustrated
in Figure 6 were changed from 150 to 50 RFU’s (relative
fluorescent units) in an attempt to detect more alleles.

When interpreting the genetic profiles, the following
guidelines were followed:

1. Alleles of a genetic profile with an RFU value of
150 to 4500 are declared true alleles [3].

2. Alleles of a genetic profile with an RFU value of
50 to 149 are declared active.

3. Alleles of a genetic profile that fall below 50
RFU’s are declared undetectable.

4. For alleles from a heterozygous individual at a
particular locus, heterozygote peak ratios are
determined by dividing the peak height of the
allele with the lowest RFU value, by the peak
height of the allele with the largest RFU value
and multiplying this value by 100 to obtain a
percentage. This value must be 3 70% for a
heterozygote to be declared. Any ratios below
70% should be interpreted with caution.

5. A sample can be considered to have originated
from a single source if 1) only one or two
alleles are present at all loci examined, and 2)
the peak height ratios of heterozygous
individuals at a locus are within the expected
range.

6. A sample can be considered to have originated

from multiple (two or more) sources if 1) more
than two alleles are present at two or more loci,

27

 

 

 

 

 

* _ “ “ 7 ‘ I ' ‘ :5 H n a" '3'! S I S P a [a m B t e rs gig;1;."iii-2::Li';:'.:.:.§" a;

 

.— «a v onto”... --—.~—..-‘ ... .~. .0..-.- .. .-

 

——flnalgsis Range

 

 

—-Size Call Range

 

 

 

 

 

E MultiComponent
——Smooth Options
C) None

Ci Light

(E) Heavu

 

 

 

 

0 Full Range 0 an Sizes
6) This Range (Data Poi nts) © This Range (Base Pairs)
Start: Min:
Stop: Max:
——Data Processing —Si2e Calling Method —
E Baseline 0 2nd Order Least Squares

0 3rd Order Least Squares
0 Cubic Spline Interpolation
(Q) Local Southern Method

0 Global Southern Method

 

 

 

 

—Peak Detection
Peak Amplitude Thresholds

 

e-

 

 

 

<81

 

'5
1:,-

 

 

Min. Peak Half Width: [_3—___| Pts

 

 

—$plit Peak Correction —

(E) None
0 GENESCAN 2500

O Left Most Peak
0 RightMost Peak

Correction Limit: Data Pts.

 

 

 

 

 

 

I Cam 1

 

 

 

 

Figure 6 - Genescan® analysis parameters.

 

 

and/or 2) the peak height ratios for
heterozygotes fall outside the expected range.

7. Stutter is an expected minor product peak that is
four base pairs, or one repeat unit, shorter than
the main allele [26]. Table 5 was used as a
guideline in determining the maximum % stutter
allowed at a particular locus. Peaks in the

stutter position that exceed the maximum %
stutter value may be designated as a true allele.

8. A complete genetic profile is declared if all 10
loci exhibit alleles with an RFU value of 150 to
4500, and the heterozygote peak ratios are
acceptable.

9. A partial genetic profile is declared if at least
two out of the ten loci exhibit alleles with an
RFU value of 150 to 4500, and the heterozygote
peak ratios are acceptable.

10. An active genetic profile is declared if 1) only
one out of the ten loci exhibits alleles with an
RFU value of 150 to 4500 and the heterozygote
peak ratios are acceptable, and/or 2) loci
exhibit alleles with an RFU value of 50 to 149.

11. An undetectable profile is declared when all
alleles fall below 50 RFU’s.

New Procedures

 

In an attempt to increase the percentage of complete
genetic profiles, new procedures, not previously utilized
by the manufacturer, were developed.

One of the new procedures developed was to add more
PCR product to the capillary electrophoresis reaction
mixture. Instead of adding 1ul of PCR product to the 25 ul

of formamide/ROX solution, 3 ul were added. Another

29

Table 5 — Stutter percentages.

 

 

 

 

 

 

 

 

 

 

 

DYE COLOR LOCUS CHROMOSOME % STUTTER*
5-FAM Blue D3S1358 3 15
5—FAM Blue VWA 12 15
S-FAM Blue FGA 4 15

JOE Green D881179 8 12

JOE Green D21S11 21 15

JOE Green D18851 18 18

NED Yellow D58818 5 12

NED Yellow [m38317 13 12

NED Yellow D78820 7 12

 

 

 

 

 

 

*Acceptable values established by Applied Biosystems [3]
and confirmed by Michigan State Police validation studies.

30

procedure developed was using Millipore microconC>
centrifugal filter devices to concentrate the PCR

product. First, the filter was washed by adding 100 ul of
TE buffer and spinning down (500 g/15 minutes). All of the
PCR product was then added to the filter and spun down

(500 g/6 minutes). The filter was removed from the tube,
inverted, placed into a new tube, and spun down (1000 g/3
minutes). This tube now contained the concentrated PCR
product. Three microliters of this concentrated product
were then added to the 25 ul of formamide/ROX solution.

Both procedures developed involve direct manipulation

of the PCR product generated from the Profiler PluS”‘kit.
Since I chose to use the reagents supplied in this kit and
the protocols established by the Michigan State Police
laboratory, manipulating the actual PCR reaction in hopes
of generating more PCR product was not an option. If it
were an option, varying reagent concentrations,
manipulating PCR cycles, and developing a nested PCR

reaction [21] would have all been explored.

Quality Control (QC)/Quality Assurance<QA)
In 1988, the Federal Bureau of Investigation (FBI)
formed the technical working group on DNA analysis

(TWGDAM). Currently known as SWGDAM (scientific working

31

group on DNA analysis), the function of this group was to
provide a forum to discuss various issues concerning
forensic DNA laboratories. Specifically, QA/QC issues were
addressed and guidelines for a quality assurance program
for DNA testing laboratories was established. In 1994,
Congress passed the DNA Identification Act which created
the DNA Advisory Board (DAB). This group, comprised of
members appointed by the Federal Bureau of Investigation,
was also created to develop standards for DNA testing and
quality assurance. Together, both groups have established
guidelines which, if followed, can assure the reliability
of forensic DNA testing methods in producing accurate and
precise results.

The American Society of Crime Laboratory Directors
(ASCLD) is an agency responsible for assessing a
laboratories compliance with guidelines established by
SWGDAM and the DAB. The Michigan Department of State
Police Crime Laboratory in Northville, Michigan, was found
to be in full compliance with all established guidelines
and has been awarded accreditation by the ASCLD Laboratory
Accreditation Board. In order to assure continuous
compliance with ASCLD guidelines, the laboratory is re-
inspected every four to five years. In regards to this

research, ASCLD standards for procedures, equipment and

32

physical plant were all followed in order to assure the
reliability of the results. Specifically, laboratory
design, validation of the testing method, technique of the
scientist, and incorporation of appropriate QC samples will
be discussed in more detail to illustrate their importance
in this particular study.

Laboratory design is critical when performing forensic
DNA analysis. In order to prevent contamination of
unamplified DNA (pre—PCR) with amplified DNA (post-PCR),
Applied Biosystems recommends designating the following
work areas: evidence handling, DNA extraction, PCR setup
and amplified DNA [3]. The first three work areas are pre—
PCR rooms while the fourth is a post-PCR room. It is
required that pre- and post- rooms are physically isolated
from one another and contain designate equipment and
supplies that never leave their assigned work area (ie. lab
coats, pipets, pipet tips, reagents, etc). Figure 7
illustrates this type of laboratory design [3]. The
Michigan State Police forensic biology unit in Northville,
Michigan, has incorporated these recommendations into their
own laboratory design.

Both the Applied Biosystems AmpFlSTRC>Profiler PlusTM

PCR Amplification Kit and the ABI PrismC>310 Genetic

33

 

EVIDENCE HANDLING

 

WORK AREA DNA EXTRACTION WORK AREA
Evidence examination Sample digestion {DNA-quandtatidn-(do-t Bio-t pr-dc-ed-ui'el ]

 

Differential lysis

Organic extraction“
microconce ntration

Stains. specimens divided
for DNA extraction

Storage oi extracted DNA samples
and AmpFISTR Profiler Plus Kit
components (except AmpFrSTR'

 

 

 

 

 

 

 

 

Chelex extraction AHQHC Ladders)
Evidence ir Midget-op; 1
photography L ------ 4
' PCR SETUP WORK AREA
Microscopy PCR reagent and DNA

sample additions

 

 

 

 

 

 

 

 

AMPLIFIED DNA WORK AREA DARK ROOM
r----—-:—f-—‘1 Photography
l DNA quantitation i DNA Thermal Cycler 480. . Developed OuantiBlot-
: (slot blot procedure) : GeneAmp’ PCR System 9600. and membrane
e ........... a GeneAmp PCR System 2400

Film Development
- Chemiluminescent
DNA quantitation Amth’STR Profiler Plus detection QuantiBlot membrane

 

 

 

 

 

 

 

 

- I - lot
(p08! 8 0t b ‘) - ABI Parser 377 and
ABI PRISM 310 instruments

 

 

 

 

 

 

 

 

 

 

 

~ Gel pouring
Storage of amplified DNA Waste disposal of amplified
samples and Allelic Ladder products
Figure 7 — Laboratory design.

34

Analyzer have been subjected to rigorous validation
studies. These studies, designed by TWGDAM and the DAB,
are necessary in order to prove the testing method reliable
validation studies on both the kit and the instrumentation.
for forensic DNA casework [12]. This re-inforces the fact
that manipulating the PCR reaction in forensic casework is
not an option. The current cycling parameters and reagent
concentrations have been established by TWGDAM and the DAB
and must be upheld. Changing any of these established
parameters and concentrations is a lengthy and time
consuming process which requires a consensus of the
forensic scientific community employing the particular
method. In addition, the Michigan State Police laboratory
has conducted their own validation studies on both the kit
and the instrumentation. As a result of these studies, the
Michigan State Police has documented specific procedures
that must be followed when using this kit and
instrumentation in forensic DNA casework. All of these
procedures were followed when conducting this research.
The technique of the scientist performing the testing
also plays a critical role in preventing contamination.
The following is a list of precautions that Applied
Biosystems would like all practicing forensic DNA analysts

to consider [3]:

35

10.

ll.

Examine and sample from pieces of evidence at a
separate time from the handling and sampling from
known samples.

Use disposable gloves at all times and change
them frequently.

Always use a clean cutting surface and clean
scissors when collecting evidence samples.

Perform DNA extraction from samples containing
high levels of DNA separately from samples
containing low levels of DNA.

Perform the DNA extraction of evidence samples at
a separate time from the DNA extraction of known
samples.

If possible, PCR reactions should be setup in a
dedicated PCR hood equip with a UV light source.

Use sterile, disposable, hydrophobic filter—
plugged pipet tips and microcentrifuge tubes.

Always change pipet tips between handling each
sample.

Store reagents as small aliquots to minimize the
number of times a given tube of reagent is
opened.

Cap all tubes before beginning the addition of
DNA. Only open the tube to which DNA is being
added.

Do not store reagents close to samples containing
high levels of DNA.

All of these recommendations were followed in order to

minimize the possibility of contamination.

Running appropriate quality control samples is also

very important in assuring that contamination is not

occurring and to guarantee the testing methods ability to

36

produce accurate and precise results. In this study,

QC samples, positive and negative controls, were introduced
into the testing method at three very distinct points of
analysis.

First, was during the extraction process. An internal
laboratory control (ILC) and a substrate control were both
extracted in parallel with research samples. The ILC has a
known DNA profile and is used by the Michigan State Police
as a positive control. The substrate control was used as a
negative control. This sample was collected randomly after
the handle was disinfected but before the next individual
came into contact with it. A negative result for this
sample assures that the disinfection process is working
properly to prevent DNA carryover from occurring.

Second, was during the PCR setup process. Again,

both a positive and negative control were setup in parallel
with research samples. AmpFlSTR®>Control DNA 9947A,

provided in the Profiler Plusm‘kit, was used as a positive
control. This control DNA has a known DNA profile, as
established by Applied Biosystems. The negative control
was PCR master mix without the addition of DNA.

Third, was during the capillary electrophoresis setup

process. An additional negative control was run in

37

 

parallel with research samples. This sample consisted of
310 master mix without the addition of PCR product.

A total of two positive controls and three negative
controls were run with every batch of samples that went
through analysis. If unexplainable discrepancies exist
with any one of the control samples, results generated from
that particular batch of research samples are deemed

invalid and need to be re-tested.

38

RESULTS AND DISCUSSION

Decontamination Testing

 

QuantiBlotC>results confirmed that the male
consistently transferred DNA to the handle of the hammer.
Since male/female mixtures were not seen in any of the DNA
profiles generated from the female samples, the

decontamination process was working effectively.

First Round of Sample Collection

 

Yield Gel Results

No visible bands were observed in any of the 32
sample lanes, which represents a concentration of DNA
less than 3.75 ng/ul (15 ng/4ul). All results

obtained from QC samples were acceptable.

QuantiBlot® Results

DNA concentrations ranged from 0.125 ng/ul to
0.03125 ng/ul, and the results generated from QC
samples were acceptable (Table 6). Most important,
the substrate control sample did not show a visible
band in its slot. For quality control purposes, this
sample was randomly collected after the handle had

been decontaminated but before the next individual

39

Table 6 - First round quantiblot results.

 

Quantiblot Results Sample
(ng of DNA/ul)

 

0.125 HH
DD*
ILC (2)

 

0.0625 G

 

0.03125

 

 

< 0.03125 A

BB*
II
Substrate Control

 

 

*The individuals who supplied these samples were chosen to
take part in the second round of sample collection.

40

 

held onto it. This was used to assure that the
decontamination process was working properly to

prevent DNA carryover from occurring.

Second Round of Sample Collection

 

Yield Gel Results

No visible bands were observed in any of the 5,
15, or 30 second sample lanes, and all results
generated from QC samples were acceptable. The known
buccal samples showed solid, bright bands in their

respective lanes.

QuantiBlot®>Results

For all 5, 15, and 30 second samples, the highest
amount of DNA recovered was 0.0625 ng/ul (Table 7),
and the known buccal samples yielded between 0.25 and
0.125 ng of DNA/ul. Results obtained from QC samples
were all acceptable. It should be noted that in both
rounds of sample collection, a relationship did not
appear to exist between the amount of DNA quantatied
and the gender of the individual who supplied the
sample.

Not only does this data show the possibility to

quantitate DNA from holds as low as 5 seconds, it also

41

Table 7 — Second round quantiblot results.

 

Quantiblot Results
(ng/ul of DNA)

Sample

Amount of DNA to
be amplified

 

0.25

FF—KNOWN
M-KNOWN
U-KNOWN

DD—KNOWN

1.0 ng*

 

0.125

AA-KNOWN
J-KNOWN
O-KNOWN
Q-KNOWN
BB—KNOWN
ILC
N-KNOWN
X-KNOWN

1.0 ng

 

0.0625

Q-30

0.625 ng

 

0.03125

FF-5,15
L—30
J—15,30
F-15,30
H-15

0.3125 ng

 

<0.03125

 

AA-5,15,30
FF-3O
L-S,15
J—5
F-5
M-5,15,30
0-5,15,30
U—5,15,30
Q—5,15
BB-S,15,30
H—5,30
DD—5,15,30
N—5,15,30
X-5,lS,30
W-5,15,30
Substrate Control

 

<0.3125 ng

 

 

* Amount required for optimum amplification to occur, as
determined by Michigan State Police validation studies.

42

 

shows that there is no apparent relationship between
handling time and the concentration of DNA recovered
from the handle. For a true proportional relationship
to exist, there should be an increase in the amount of
DNA recovered from the 5 second hold up to the 30
second hold. The data generated in this study does
not show this to exist, as one might expect.

For example, samples FF—5 and FF-15 each gave
0.03125 ng of DNA/ul but FF—30 gave < 0.03125 ng of
DNA/ul. Also, sample H—5 gave < 0.03125 ng of DNA/ul,
H-15 went up to 0.03125 ng of DNA/ul, and H-30 dropped
back down to < 0.03125 ng of DNA/ul. Data from the
initial 2 minute holds also plays a part in evaluating
this relationship. After 2 minutes, individual Q
transferred < 0.03125 ng of DNA/ul to the handle.

At a later date, a sample was collected from
individual Q after only 30 seconds. From this sample,
0.0625 ng of DNA/ul were recovered.

Since time does not seem to be related to the
amount of DNA recovered, the condition of the test
subjects hands appears to be an important factor. DNA
from skin cells shed directly from the hands
(epithelial cells), can contribute to the DNA

recovered from the surface of the handle. One

43

previous study suggested that DNA transfer is highly
dependent on the individual handling the object [14].
Those that shed skin cells more readily than others
were categorized as “sloughers”. Natural human
variation can account for one being categorized as a
“slougher” or a “non-slougher. For example, those who
naturally have dryer hands might shed skin cells at
different rates than those who do not.

Cells shed directly from the surface of the hands
can contribute to the amount of DNA transferred to the
handle, but it does not necessarily represent the
entire amount. DNA transferred to the hands from
other parts of the body can also contribute. Before
coming in to contact with the hammer, the subject
could have coughed or sneezed into their hands,
scratched their head, rubbed their eyes, or touched
any other part of their body that contained large
amounts of nucleated cells. Each of these instances
can transfer many nucleated cells to the hands which
in turn could be transferred to the handle. In this
particular study, this was a possibility because the
hands of the individuals were not controlled in any
way prior to coming into contact with the handle.

To get a true representation of primary DNA

44

transfer occurring in the sample population, it is
essential that the hands not be controlled. In many
instances, violent criminal activity is an extremely
random event. Controlling the hands would not produce
“true—to—life” results, thus compromising the
significance of the results. This lack of control
over the individuals hands, along with natural human
variation, is what accounts for the reason why some
samples were able to be quantitated while others were

not.

STR Results

All known DNA profiles generated from the buccal
samples were complete and consistent with originating
from a single source (Table 8).

Table 9 shows the genetic profiles generated from
each 5, 15 and 30 second sample. Out of the 45 total
samples, alleles were detected in 44. This shows that
primary DNA transfer to this type of surface is very
common in this particular population.

After each profile was subjected to the
interpretation guidelines listed previously, its DNA
profile type was determined (Table 10). The majority

of the samples, 20 out of the 45, produced partial DNA

45

cocmflanwumo >Hmsofl>oud one: moaflwoud «zo czocxae

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

«H.0H Ha.oa Ha.m ma.ea om.om ea.ma x.x N.N~.ma oa.ea ea.oa eem execs
oa.m mH.HH Ha.oa aa.ma N.om.om mH.HH x.x am.mm ma.ea oa.ma are means
ma.a NH.HH ma.~a ma.ma om.mm Ha.m x.x mm.mm ma.ma ea.oa seq geese
HH.HH Hates ma.aa ma.ma om.em ea.ea x.x em.mm ma.ma oa.ma eee seeps
Na.ma HH.HH NH.HH ea.ma om.mm ma.aa ».x em.em ma.ma oa.oa mm paces
Ha.oa Na.m ma.aa ma.ma Hm.om ma.ma x.x mm.em ea.aa ma.ma o seeps
Ha.m Ha.m NH.HH Hm.wa m.am.mm ma.ma ».x om.am ea.ma ea.ea 8 cases
NH.NH NH.HH ma.a ma.ma Hm.em ea.m x.x om.em ma.ma ea.ma o esoem
aa.ea NH.HH ma.aa ma.ma om.om aH.HH x.x mm.mm aa.ea oa.oa .2 seems
HH.HH NH.HH aa.aa ea.ma mm.mm ma.oa x.x Hm.am aa.ea oa.ea a axons
NH.HH NH.HH ma.oa ma.ma em.em ea.ma w.x em.am ma.ma ma.ma ea seeps
NH.oa Ha.m HH.HH ea.ea om.mm ma.ea ».x am.mm ma.ea oa.ma we eeoem
oa.oa Ha.oa ma.a mH.NH am.mm ma.ea s.x mm.mm matea ma.ma e seeps
oa.a NH.HH NH.Ha ea.ma N.om.mm ma.oa ».x mm.ma ea.oa ma.oa x macaw
«H.0H HH.HH NH.HH ea.ma om.mm ma.aa x.x om.mm ma.ma oa.oa 2 seems
oa.e NH.HH HH.HH ea.ma am.mm aa.ma w.x mm.am ea.oa oa.ma on ozone
oummpo 5339 333 Romeo 333 2.3me an 4mm 65 $3me 3 395m

 

 

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46

nauseouonm 3w: on pomOQXo mums memEmm paom
ouduxfle n no oocopfl>o pesosm moHQEmm ee

Am.aun mea-omv

wmamaam o>fiuo¢

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HH.HH NH.HH ea:aa ea.ma mm.mm ma.oa x.x Hm.am ma.ea oa.ea mH-e
‘HH «Eff: awataaa 4¢HKNH mmtmm gmafoa “TN HNtHN «mHHH wail” mih.
ema.eaa ema.eaa ema.eoa mH.mH amm.aem ea.ma w.x em.em ma.ma mH.mH om-mm
mataa mataa mated malma mm.em ea.ma w.x em.em matma ma.mH ma-mm
NH.HH NH.HH ma.oa ma.ma mm.em «Hima w.x em.em ma.ma mH.mH m-mm
‘OH 239% HHJH <bHc¢mH «‘omlmm mHth Nix agemfmm ¢mdc<ha 3”th amt:
eea ew.ex ems ewe ma-<<
eHH 2mm eve ew.«x eea eeofl.ema mrae
oa.ea HH.OH ma.m mHtNH mm.mm ma.ea w.x mm.mm ma.ma ma.oa om-3
40H aHHl‘OH 4mH.4m eemat‘ma mmtmm matva w.x aamm.mm «mat<ma mH.mH mar:
eoa eHH eema.em ‘ma amm.4mm ema.eea >.x emm.emm «matema ema.ema m-z
eoa.em amateaa NH.HH eatama eem.om.mm ema_oa w.x mm.ma ma.oa ma.ma om-x
eoa ema ema.xaa exea.ama em.om.emm MH.OH w.x emm.4ma ema.eoa ma.ma mars
«oatem ema.eHH ema.eHH eeeateMH «m.om.mm mH.oa w.x mm.ma aema.oa ma_oa m-x
«somiz
awe mH-z
semiz
camataaa aHtHH 4MH «mm.amm eava_ama M.x ‘Hm matma ema.‘ma omiDQ
oatm NH.HH HH.HH ea.ma mm.mm eH.mH w.x mm.am mH.mH ma.ma mH-oo
oa.m mH.HH HH.HH eatma mm.mm eatma w.x mm.am ma.ma matmfl m-oo
enemas bananan mammmn Hmmman Hamaaa meaammo MN «on ¢z> mmmammn nH odesom

 

 

 

 

 

 

 

 

 

 

 

 

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47

moedcououm 30c 00 pomomxm onoz mmHQEMm paom
annexes a mo oommcfi>o posocm oHQEmm «e
Am.DwH m¢Hiomv

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_ «tomim
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“teal aaaw eHH.eoa ama em.om.xom oema.aa x.x aem.emm eama.eea oa.ma m-m
aeomrq
¢<matam <mHt<HH MH.NH ema.<mH om.emm Hatm x.x e<mm.mm mHYmH mH.mH mHiq
«smug
aaomim
HH.HH HH.HH ‘mH.HH ma.ma oH.mH ma-m
:HH HHtHH amataa a:ma.amfl mH.mH mim
ewe HH.HH NH.HH ma.ma oH.oH om-mm
ama HH.HH ma.eaa ma.ma oa.oa ma-mm
ewe eHH eama.‘HH ma.ma mates m-mm
aaa.oa mH.HH ea.ea ma.ma em-o
«Hacaoa mHiO
aaa.eoa m-o
. om-p
mH-s
a. .. . m-o
. ma om-o
ewe ma-o
,- n_asfie m-o
eeom-z
‘HHtaoH ama.‘aa om.om vH.HH x.x amm.~mm emat4ea mH.wH mar:
enemas bammmao wammmn Hmmman Hamaao meaammn we 4cm 43> mmmammn nH oaaaom
.Ae_oeoov m oanme

48

Table 10 — 5,15,and 30 second profile types.

 

 

 

 

 

 

 

 

 

Type of DNA profile 5 seconds 15 seconds 30 seconds
Complete DD-S FF—15 W-30
FF-S H-15 J-3O
DD—15
J-15
Partial X-S X—lS DD—3O
J—S W-lS X-30
Q—5 M—15 AA—3O
BB—5 Q—15 FF—30
F-5 BB-lS O-30
H—5 F-15 Q-30
L—lS BB-30
Active W—S N—15 U-3O
AA-S AA-15
M—5 O-15
0—5 U—15
Uhdetectable U—5
Mixture N-S
L-5

 

 

 

 

 

49

profiles. Of the remaining 25 samples, nine gave
active profiles, eight gave complete profiles, one
profile was undetectable and seven profiles showed
evidence of a mixture. In mixtures, alleles belonging
to the primary handler are present, but additional
alleles, not belonging to the handler, are also
detected. Again, it should be noted that

a relationship did not appear to exist between the
type of DNA profile generated and the gender of the
individual who supplied the sample.

These results are extremely significant when
evaluating the sensitivity of the testing method.
With one exception, all of the extracts contained DNA
concentrations 5 0.03125 ng/ul. Therefore, the DNA
amounts introduced into the amplification reactions
were 5 0.3125 ng (312.5 pg). This is much lower than
the 1.0-1.25 ng recommended by the manufacturer [3].
Generating complete and partial DNA profiles from
samples containing such low copy numbers (LCN) of
target DNA, as this study did, illustrates the extreme
sensitivity of PCR/STR technology.

As the DNA profiles were being interpreted, two
issues surfaced which need to be addressed. The

first issue is the occurrence of heterozygote peak

50

imbalances. The STR interpretation guidelines, based
on results generated from Michigan State Police
validation studies, state that the heterozygote peak
ratios must be 3 70% for a heterozygote to be declared
at a particular locus. When looking at the 38
samples that did not show evidence of a mixture,
imbalances (ratios <70%) were seen in 45% of them. Of
this 45%, 27 separate instances occurred with the
majority seen at the VWA and D18S51 loci.

Of the 27 instances, only one instance involved
heterozygote alleles 3 150 RFU’s. The Michigan State
Police interpretation guidelines states that true
alleles are declared if they fall between 150 and 4500
RFU’s [3]. In this study, a new guideline declared
active alleles if they fell between 50 and 150 RFU’s.
Without this guideline in place, imbalances would have
occurred in only 3% of the samples.

Explanations as to why these imbalances occurred
include primer binding site mutations and low copy
number DNA [25]. Given that this study directly
involved the analysis of low copy number DNA, this
appears to be the reason why imbalances occurred.

With low levels of input DNA, it is possible that two

alleles of a heterozygote will amplify unequally.

51

This phenomena is known as stochastic fluctuation
[13].

It should be noted that not all laboratories
use the same heterozygote peak ratio guideline. In
studies conducted by the Federal Bureau of
Investigation, ratios 3 60% were deemed acceptable
[18]. If this guideline were used to interpret the
DNA profiles generated in this study, only 16% of the
samples would have shown imbalances.

The second issue involves the seven samples (16%)
that showed evidence of a mixture. No more than three
loci in any one sample showed additional alleles, and
no more than one additional allele was seen per loci.
Again, it should be recognized that if the active
allele guideline was not incorporated into this study,
only one sample would have showed evidence of a
mixture.

Previous studies have suggested that when dealing
with low copy number DNA, mixtures are commonly
encountered [8]. There are two explanations as to
why the mixtures could have occurred. One is DNA
contamination and the other is artifacts produced by
the electrophoretic and/or PCR system itself. In

regards to DNA contamination, heterozygote peak

52

imbalances at the Amelogenin loci (XzY) are typical of
a male/female mixtures [9]. Since this imbalance was
seen in three of the mixed samples, DNA carryover,
laboratory contamination and secondary transfer all
need to considered as possible sources of DNA
contamination.

If the procedure explained earlier to
decontaminate the handle of the hammer was not working
efficiently, this would cause DNA carryover to occur.
This would result in a mixture of alleles belonging to
the primary handler and the individual who handled the
hammer just before. Since the decontamination process
was extensively tested prior to beginning the study,
DNA carryover does not seem to be the likely reason as
to why mixtures were seen.

Research conducted by Peter Gill of the Forensic
Science Service in the United Kingdom has shown that
low copy number DNA amplification is prone to sporadic
contamination [8]. Just as in this study, strict
QA/QC guidelines were followed in order to prevent
contamination from occurring, but additional alleles
were still detected [8,11]. Possible sources of

contamination include extraneous DNA, cross

53

contamination from other samples processed in the
laboratory and plastic-ware contamination by the
manufacturer [8]. In Gills research, contaminants
were typically associated with low molecular weight
loci which included Amelogenin, D38158 and D8S1179
[11]. In this study, 67% of the additional alleles
were seen at these loci.

The occurrence of a secondary transfer is another
possible source of DNA contamination [16,24]. As
explained earlier, the hands of the individuals were
not controlled in any way prior to coming into contact
with the handle of the hammer. Therefore, the
mixtures seen in this study offer proof that it could
be possible for one individual to transfer another
individuals’ DNA to an object and have it be detected.
When explained in terms of criminal activity, it could
be possible for a criminal to transfer an innocent
person’s DNA onto a weapon of this type, thus
connecting him/her to the crime. More extensive
research needs to be conducted in this area in order
to prove this possibility to be true.

Artifacts produced by the electrophoretic and/or
PCR system itself is the second explanation as to

why the mixtures could have occurred. One type of

54

artifact is stutter. Stutter peaks are one repeat
unit shorter than the main allele and are caused by
slippage of the DNA polymerase enzyme during PCR [26].
Studies conducted by Applied Biosystems and confirmed
by the Michigan State Police showed that stutter peak
heights are typically 12% to 18% of the main allele.
This range exists because different loci show
different intensities of stutter. When low copy
numbers of DNA are introduced into the PCR reaction,
stutter peaks can actually be equivalent to the size
of the main allele [9]. In this study, four samples
contained additional alleles that were in the stutter
position.

Non-specific artifacts can also be produced by
the electrophoretic and/or PCR system. This type of
artifact occurs as a result of non—specific priming
from fragments of bacterial or degraded human DNA
[9,10].

The results of Peter Gill’s studies show that
appropriate guidelines need to be established for
interpreting DNA profiles generated from low copy
number DNA [8,11]. As recommended by Gill, an allele
should only be reported if a duplicate result can be

obtained from two or more replicate samples [11].

55

Only 4 out of 1225 comparisons showed the same
spurious allele when replicates were compared. This
“duplication guideline” was applied to a statistical
theory which proved this to be a reasonable approach
to interpreting low copy number DNA profiles.
Unfortunately, the results of Gill’s studies were not
acquired until after the research had been completed
in this study. Otherwise, an honest attempt to
replicate all samples showing evidence of a mixture
would have been made.

Tables 11 and 12 show genetic profiles generated
from 15 samples after exposure to the two newly
developed procedures. Profile types were determined
and compared to each other, and to those generated
from the samples exposed to the original procedure
(Table 13).

When using 3 ul of PCR product in the 310
reaction, two active profiles became partial, while
the remaining four showed increased activity but
remained active. One partial profile became
complete, while eight showed increased activity but
remained partial. Two out of the nine partial
profiles would have been declared complete if

heterozygote peak ratios would have fallen within the

56

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ma.ma Hm.om ma.ma x.x mm.em ea.ea ma.ma ma-o
. eema.ama <w.x aom.¢am aea eea.<ea om-o
eama.ma ew.x Hm «eea.ma reea.ea ma-o
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ama eeaa.am HH.HH eaea.aoa e40m.wm «w.x eema.ea oa.ma m-¢¢
ea.oa HH.OH ma.m ema.ma mm.wm ma.ea ».x eH.oH ama.oa ma-z
eaoa.em ma.¢aa NH.HH eeea.ma m.om.mm ma.ea w.x ema.oa ma.oa ma-x
oa.m NH.HH NH.HH m.om.mm ma.oa w.x ema.oa ma.oa m-x
eaa «He 4 .:.. gee x.x «ma oa.oa ma-z
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57

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NH.HH ma.aa ma.oa matma mmtem ea.ma wtx em.em ma.ma ma.ma om-mm
ma.oa Ha.m HH_HH halos om.mm ma.ea w.x em.mm ma_ea oa.ma om-4¢
emieoH «Ha.m HH.HA ea.oa eom.mm emH.eH w.x em.mm ema.ea eoa.ma m-<<
oa.oa Ha.oa ma.m ema.ma mm.wm ma.ea w.x emm.mm wa.oa ema.oa ma-z
oa.m NH.HH NH.HH eeatma m.om.mm ma.ea w.x mm.ma ema.oa mH.mH ma-x
ma.aa NH.HH ea.ma m.om.mm ma.ea w.x mm.ma «wa.oa ma.ma m-x
r. a .3 Ha enigma :omtamm Jami”; xtx «mmlemm «ma matma maiz
ommmeo hammman memmmo Hmmmao HHmHNn meaammn ex «on ¢z> mmmammn nH madamm

 

 

.DUSUOMQ mum commencwucoo wo dam i We canoe.

58

Table 13 — New procedure profile types.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sample ID Quantiblot Original 3ul of 3ul of
results Protocol amplified concentrated

(ng of (lul of product amplified

DNA/ul) amplified product

product)

N-lS < 0.03125 Active Partial Partial
AA-S < 0.03125 Active Partial Partial
M-S < 0.03125 Active Active Partial
O-5 < 0.03125 Active Active Mixture
0-15 < 0.03125 Active Active Mixture
U-30 < 0.03125 Active Active Mixture
Q-15 < 0.03125 Partial Complete Complete
X-S < 0.03125 Partial Partial* Partia1*
W-IS < 0.03125 Partial Partial* Partial*
X-15 < 0.03125 Partial Partial Partia1*
AA—30 < 0.03125 Partial Partial Complete
BB-S < 0.03125 Partial Partial Partial*
L-IS < 0.03125 Partial Partial Complete
FF-30 < 0.03125 Partial Partial Complete
J_5 < 0.03125 Partial Partial Complete

 

 

*All 10 loc1

70%.

59

L exhibited alleles with an RFU value of 150 to
4500, but the heterozygote peak ratios at some loci were below

 

acceptable range.

Heterozygote peak imbalances were seen in 60% of
the samples. Of this 60%, 20 separate instances
occurred with the majority seen at the VWA, D21811 and
D18S51 loci. In comparison to the same samples from
the original procedure (1 ul of PCR product), the same
nine samples showed imbalances but six more instances
were seen. Originally, no more than two loci in any
one sample showed imbalances. Adding 3 ul of PCR
product generated DNA profiles with up to five loci
exhibiting imbalances.

When comparing DNA profiles generated from the
original procedure to those generated in this
procedure, two major differences were seen. One
is the peak heights of the alleles showing
imbalances. Of the original 15 samples, only active
alleles (50-149 RFU’s) were involved. When more PCR
product was introduced into the 310 reaction, 25% of
the instances involved alleles with peak heights in
the acceptable range (150-4500 RFU’s). Another
difference is the severity of the imbalances.
Originally, the majority of the imbalances involved
alleles with peak height ratios that fell between 60%

and 70%. With this procedure, the majority of the

60

ratios fell between 40% and 60%.

When using 3ul of concentrated PCR product in the
310 reaction, three out of the six active profiles
became partial, but the remaining three showed
evidence of a mixture. Five partial profiles became
complete, while the remaining four would have been
declared complete if the heterozygote peak ratios had
fallen within the acceptable range.

Heterozygote peak imbalances were seen in 58% of
the samples that did not show evidence of a mixture.
Of this 58%, 16 separate instances occurred with the
majority seen at the VWA and D18851 loci. Up to six
loci in one sample showed imbalances and one sample
that did not exhibit imbalances with the original
procedure showed imbalances at two loci. The majority
of the peak height ratios fell between 50% and 70%,
and 75% of the instances involved alleles with peak
heights in the acceptable range. When compared to the
3 ul of PCR product that was not concentrated, the
ratio imbalances were less severe but the percentage
of instances involving peak heights in the acceptable
range increased by 50%.

Adding 3 ul of concentrated PCR product to the

310 reaction generated three DNA profiles that showed

61

evidence of a mixture. A total of 32 additional
alleles were seen. Up to eight loci in one sample
exhibited additional alleles and up to four additional
alleles were seen at a single locus. Of the alleles
belonging to the primary handler, 84% exhibited peak
heights within the acceptable range. In contrast,
alleles not belonging to the primary handler were in
the acceptable range only 16% of the time.

Both procedures were successful in increasing the
ability to detect alleles, but adding 3ul of
concentrated PCR product to the 310 reaction generated
more complete profiles. Unfortunately, results from
both procedures need to be interpreted with extreme
caution. In regards to the original 15 samples, all
heterozygote peak ratio imbalances involved active
alleles and mixtures were not seen. The new
procedures produced true alleles (150—4500 RFU’s) with
imbalances and three samples showed evidence of a
mixture. Explanations as to why imbalances and mixed
samples occurred are the same as those explained
earlier.

In addition to the two procedures developed to
increase allele detection, interpretation guidelines

established for low copy number DNA analysis could

62

have been implemented into this study and evaluated as
a more reliable method [8, 9, 10, 11]. Unfortunately,
since the results of these studies were acquired after
the research was completed, the guidelines could not
be used to interpret DNA profiles. This is due to the
fact that duplicates were not run on samples

exhibiting imbalances and/or evidence of a mixture.

QA/AC Results
Acceptable results were obtained from all quality
control samples. This assures that all results

generated from test samples are accurate.

63

CONCLUS IONS

In a population of Caucasian males and females over
the age of 15, the following conclusions can be made

regarding primary DNA transfer to an unfinished wood

surface:
1. Primary transfer is extremely common.
2. No apparent relationship exists between handling time

and the amount of DNA recovered from the substrate.

3. Complete DNA profiles can be generated from holds as
low as 5 seconds where < 0.3125ng of DNA is amplified.

4. Adding 3ul of PCR product to the 310 reaction slightly
increases the ability to detect alleles, but
imbalances in heterozygote peak ratios of true alleles
can occur.

5. Adding 3ul of concentrated PCR product to the 310
reaction significantly increases the ability to detect
alleles, but imbalances in heterozygote peak ratios of
true alleles can occur.

64

SUGGESTIONS FOR FURTHER RESEARCH

Since DNA transfer has only recently become a topic of
interest, conducting further research is going to be
critical in fully understanding its forensic significance.
It is crucial that this significance be determined in order
to assess just how much of an impact it will have on the
criminal justice system. In parallel with this study, the
following is a list of variables that could be changed in
order to gain additional knowledge in the area of DNA
transfer:

1. Use a test object made of a different substrate
(plastic, glass, finished wood, etc.).

2. After the object is handled for the allotted time,
expose it to a variety of storage conditions before
collecting the sample in order to determine if the
transferred DNA will be able to withstand its
environment.

3. Allow the test subjects hands to come into contact
with another individual before handling the object.
By doing this, the ability to detect secondary
transfers can be further studied.

4. Attempt to recover the amount of DNA required for

optimum amplification by using Millipore microconC>
centrifugal filter devices to concentrate the DNA

extract. This could eliminate the need to concentrate
the PCR product and allow for less heterozygote peak
imbalances.

65

REFERENCES

66

[l]

[2]

[3]

[4]

[6]

[8]

[9]

REFERENCES

Access Excellence. Desktop Electrophoresis Lab.
[Online] Available
http://www.accessexcellence.org/AE/AEPC/WWC/l993/movin
g.html, September 18, 2000.

Agarose gel fractionation of RSA l digested genomic
DNA. [Online Image] Available
http://www.biotech.ufl.edu/~etcl/mc_picture_gallery.ht
m, December 31, 2001.

Applied Biosystems Corporation. AmpFlSTRC>Profiler Plus
User’s manuals. Foster City, CA 2000.

Applied Biosystems Corporation. Introduction to
Microsatellite Analysis. [Online] Available
http://www.appliedbiosystems.com/molecularbiology/abou
t/dna/377/377a2a.html, September 18, 2000.

Applied Biosystems Corporation. QuantiBloU® Human DNA
Quantitation Kit package insert. Foster City, CA 1996.

Capillary Electrophoresis Theory and Background.
[Online Image] Available
http://www.ceandcec.com/cetheory.htm, December 18,
2000.

Fereday, L. Technology Development: DNA from
Fingerprints. [Online] Available
http://register.aspensys.com/nij/dnamtgtrans6/trans-
i.html, August 31, 2000.

Gill, P. “Application of Low Copy Number DNA
Profiling.” Croatian Medical Journal 42 (2001): 229-
232.

Gill, P., Sparkes, 8., Clayton, T. M., Whittaker, J.,
Urquhart, A., Buckleton. “Interpretation of Mixtures

Based on Peak Area — Identification of Genetic
Anomalies, Stutters and Other Artefacts.” [Online]
Available

http://www.promega.com/geneticidproc/eusymp2proc/16.pd
f, February 1, 2002.

67

[10]

[12]

[l3]

[14]

[15]

[l6]

[17]

[18]

[19]

Gill, P., Sparkes, R., Kimpton, C. “Development of
guidelines to designate alleles using an STR multiplex
system.” Forensic Science International 89 (1997):
185-197.

Gill, P., Whitaker, J., Flaxman, C., Brown, N.,
Buckleton, J. “An investigation of the rigor of
interpretation rules for STRs derived from less than
100 pg of DN .” Forensic Science International 112
(2000): 17-40.

Holt, C. L., Bunocristiani, M., Wallin, J. M., Nguyen,
T., Lazaruk, K. D., Walsh, P. S. “TWGDAM Validation of
AmpFlSTRm PCR Kits for Forensic DNA Casework.” Journal
of Forensic Sciences 47 (2002): 66-96.

Inman, K. & Rudin, N. An Introduction to Forensic DNA
Analysis. Boca Raton, Fl: CRC Press, 1997.

Kisilevsky, A. & Wickenheiser, R.A. (in press). “DNA
PCR Profiling of Skin Cells Transferred Through
Handling.” Canadian Society of Forensic Science.

Kobilinsky, L. “Deoxyribonucleic Acid Structure and
Function.” Forensic Science Handbook, VOlume III. Ed.
R. Saferstein. Englewood Cliffs, NJ: Prentice-Hall,
1993, pp. 287-357.

Ladd, C., Adamowicz, M. S., Bourke, M. T.,
Scherczinger, C. A. & Lee, H. C. “A Systematic
Analysis of Secondary DNA Transfer.” Journal of
Forensic Sciences 44 (1999): 1270-1272.

Lambda DNA/HindIII Marker, 2. [Online Image] Available
http://www.fermentas.com/catalog/markers/marksmo101.ht
m, December 30, 2001.

Moretti, T. R., Baumstark, A. L., Defenbaugh, D. A.,
Keys, K. M., Smerick, J. B., Budowle, B. “Validation
of Short Tandem Repeats (STRs) for Forensic Usage:
Performance Testing of Fluroescent Multiplex STR
Systems and Analysis of Authentic and Simulated
Forensic Samples.” JOurnal of Forensic Sciences 46
(2001): 647-660.

Structure of DNA. [Online Image] Available

http://www.accessexcellence.com/AB/GG/structure.html,
December, 18 2000.

68

[20]

[21]

[22]

[23]

[24]

[25]

[26]

[27]

[28]

Sweet, D., Lorente, M., Lorente, J. A., Valenzuela, A.
& Villanueva, E. “Improved Method to Recover Saliva
from Human Skin: The Double Swab Technique.” JOurnal
of Forensic Sciences 42 (1997): 320-322.

Strom, C.M, Rechitky, S. “Use of nexted PCR to
identify charred human remains and minute amounts of
blood.” JOurnal of Forensic Sciences 43 (1998): 696-
700.

The Polymerase Chain Reaction. [Online Image]
Available
http://www.acessexcellence.org/AB/GG/polymerase.html,
December 20, 2000.

Van Hoofstat, D. E. O., Deforce, D. L. D., De Pauw, I.
P. H. & Van den Eckhout, E. G. “DNA Typing of
Fingerprints using Capillary Electrophoresis: Effect
of Dactyloscopic Powders.” Electrophoresis 20 (1999):
2870-2876 .

Van Oorschot, R. A. H. & Jones, M. K. “DNA
Fingerprints from Fingerprints.” Nature 387 (1997):
767.

Wallin, J. M., Holt, C. L., Lazaruk, K. D., Nguyen, T.
H., Walsh, P. S. “Constructing Universal Multiplex
PCR Systems for Comparative Genotyping.” JOurnal of
Forensic Sciences 47 (2002): 52-65.

Walsh, P.S., Fildes, N.J. and Reynolds, R. “Sequence
analysis and characterization of stutter products at
the tetranucleotide repeat locus vWA.” NUcleic Acids
Res. 24 (1996): 2807-2812.

Wickenheiser, R. A. & Challoner, C. M. (in press).
“Suspect DNA Profiles Obtained from the Handles of
Weapons Recovered at Crime Scenes." lOw'Annual
Symposium on HUman Identification. Promega, Lake Buena
Vista, Florida, 1999.

Wickenheiser, R. A., Roney, J. M., Hummel, K. H. J.,
Szakacs, N. A., MacMillan, C. E., Kuperus, W. R.,
Walker, T. J., Hrycak, T. L., Reader, L. J. V.,
Fenske, B. A., Hanniman, J. L., Faris, J. S., Lett, C.
M., DeGouffe, M. J., Golin, M. & Jobin, R. M. “Unusual
Exhibit Material Yielding Successful DNA Profiles

69

[29]

Using PCR Str Typing.” 10Ch Annual Symposium on HUman
Identification. Promega, Lake Buena Vista, Florida,
1999, Poster Presentation.

Wiegand, P. & Kleiber, M. “DNA Typing of Epithelial

Cells after Strangulation.” International JOurnal of
Legal Medicine 110 (1997): 181-183.

70

APPENDICES

71

APPENDIX A

Electropherograms generated from decontamination testing.

72

ABI Thesis data - contamination study
licensed to Michigan State Police. DNA Unit Genotypem 2.1

r'lrlvrrfi'hrirrrr'riir“IVfrrrr'trwflrt
180 200 220 240 250 280 300 320

 

r—ITTTUTU"? 'VIVVrl

100 120 140 160

III‘|II.ICIICII'OII|III'IOI'IIC‘DIJ'III'OOI’IIU‘III

too 120 :40 160 180 200 220 240 260 280 300 320
Aii-P (C) 7am P (o)

 

A1 14’ (C)
800

 

   

oo
zoo
Ail-P (o) 7 Yellow P to)
too
. o
B m 51:1. If}
ED El E8
[Hi Ell
HE
Mi-P to) 7 M P (C)
E800
800
L400
[200

 

 

 

 

 

73

 

east

Thesis data - contamination chock,
Licensed to Michigan State Police, DNA Unit

Genotyporo 2.1

 

Ifiw

100

I | I I I
1 oo
A9-L (0)

A94- (0)

AQ-L (C)

NH- (C)

r
120

I
120

 

‘_T I I I I—T—I—I' l U I

140 160
I I I I I I I |
140 160
6 Blue L (C)

sateen L (C)

s‘fiflbw Lie)
FE
6Red L(C)

180 200 220 240

180 200 220 240

 

 

T I' I r U I l I Y I I’ I I ‘I’ r, r I I

f
280 300

280 300

' III—l l

200
1 50
hIOO
~50

 

. 300
zoo
Lioo

 

20

 

 

 

 

74

Thesis data - contamination check
Licensed to Michigan State Police. DNA Unit Genotypew 2.1

Y—rT—IIT—IWII’IIII'IjI'fi'IIT'rIII'jjIITIj—ll‘ljfiriIll—IWUI[TU—l—I
0 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360

0 80 100 I20 I40 160 180 200 220 240 260 280 300 320 340 360

A7-HiC) 5 Blue HIC)

4o
30
20
to

A7-H(C) 5 Green H(C)

20

EE

AT-HlC) 5 Yellow HICI

 

‘— +

 

 

 

75

APPENDIX B

Electropherograms generated from known DNA samples.

76

ABIA,
PRISM

Thesis date - Group 1

 

 

 

 

 

 

 

Licensed to Michigan State Police, DNA Unit GenotyporO 2.1
"IIIYTTIfirlI'Y—T‘FIIII‘U—I letjirfY'TrIrj‘l‘rYIIY—I—IT
F0 100 120 140 160 180 200 220 240 260 280 300
'III’III’III’IIIIIII'IIIIIII‘III'lll'lIIIIII|I
0 100 120 140 160 180 200 220 240 260 280 300
BLKNOWNDD 98in. KNOWNDD
3000
2000
l 1000
is [E El
2689 HE] m
[E 23
2317 m
B4—KNOWNDD 96mm KNOWNDO
r2000

-1000

 

 

 

 

 

“KNOWN DD

i

am

9 Red KNOWN DD

111

 

 

 

 

F
~2000
h
~1000
L

 

900
00
300

 

77

 

%_ Thesis data - Group 1
PR Licensed to Michigan State Police. DNA Unit Gonotypoio 2,1

VITfI’T-ITITrYTIIrIrI—TT—IrTj“[IITIVFII’T—IT‘T‘ITIVIIUIYI

100 120 140 160 180 200 220 240 260 280 300 320 340

 

lll'lll'llll'll IIIII'I II'III'III'Ill’ill'llO'IIIIIII'
100 120 I40 150 180 200 220 240 250 280 300 320 340
BIZ-KNOWNN 1380.10 KNOWNN

4000
3000

2000
1000

m m a": as
was as:

BiaKNOWN N 13 Green KNOWN N

 

 

 

 

 

 

£4000
-2000
I it .AJL ’
8 i3 re
5762 1866 [SE
[B

 

BIZ-KNOWN N 13 YOIIOW KNOWN N
tISOO

~1000
I I -500

 

 

 

 

IE
39
Biz-KNOWN N
900
600
300

 

 

 

78

 

ABi.A. Thesis data - Group 1 (re-run)

PRISM Licensed to Lynn. Helton, Michigan State Police GOHOWQIG 2. 1

TtIurrrIIIITr1r1{firjYWrYj—TITfifiY1‘TVI—[rltl’I‘fI—‘fiY—I‘tr“:
100 120 140 160 180 200 220 240 260 280 300 320 340 350

 

llllll'Ill'llIllllllll'IIl‘lIlllllllilllll'lII‘lll‘lII'l
I00 I20 I40 I60 I80 200 '220 240 250 280 300

320 340 360
CIT-KNOWN X 7 Blue KNOWN X

2000
1000

 

 

 

 

 

3000
2000
1000

 

1000
500

800
000

00
200

 

 

 

 

 

 

79

ABIA
PRISM

Thesis doll - Group 1 (rs-run)

 

 

 

 

 

 

 

tat—mower w

 

 

 

A I
14
IE
“I”

12 Yellow KNOWN W

11

is
m

12 Rod KNOWN w

 

 

 

. Licensed to Lynn. Helton, Michigan State Police Genotypuc 2,1
lfi'llir'r'II—lrrj'llfIIT—TT—IIU‘IIIj—Yrilfi‘I'TIfI—IIIrIII
100 120 140 160 180 200 220 240 260 280 300 320
II‘IDIIIIIIIOI‘IOIIIIt'll!’lllllll'lIOIIll‘lllll
100 120 140 180 180 200 220 240 260 280 300 320
04—KNOWN W 12 Blue KNOWN W
2000
. RH 1000
E. m V a:
an m. m
18 18 EB
[Em
D4—KNOWN W 12 Green KNOWN W

1000
500

800

00
200

 

80

 

ASIA Thesis data - Group 1 (re-run)
PRiSM

Licensed (0 Lynne Heiton. Michigan State Poiice GenotyperO 2_1
I VT 1 " I I I WI 1 I‘j I fT r I 1"] T I f l T I l I l I T—r 1—I fi‘ r T‘I— I ‘I r_l T I
100 120 140 160 180 200 220 240 260 280 300 320

 

I I ‘ I I I ' I I I I I I I | I I I ‘ I I «I ‘ I I I I I I I ‘ I I I ‘ I I I ' I I I l I I I I I
100 120 140 180 180 200 220 240 260 280 300 320
DIZoKNOWN AA 19 Blue KNOWN AA

 

 
 
 

 
   
      
   

 

 

   

3000
2000
A,jkl 1000
mg a; ’
EIIEi 115i!
In a
El!!!
D12-KNOWN AA 19 Green KNOWN M
2000
i h 1000
mg 33
EE!!! [£11m
IE 30
2 379 m
BIZ-KNOWN AA 19 Yellow KNOWN AA
P3000
Lzooo

 

 

 

;1000
*__J\.JL__ '
Ill Ell]!
liflfli IIEIM
IIIE!
01%KNCWWHAA waned KNOWNdAA

900
600
300

 

 

 

 

ABIA,
PRISM

YTIITTYT—rr'IrI’Y‘rI'l

Thesis data - Group 2
Lcensed to Michigan State Police. DNA Unit

Genotyper‘b 2. 1

 

200 220

C1-FF-KNOWN 14 Gwen FF-KNOWN

ii

IIII'III'III|IIIIIIIIDIIIIIIIIIIIIII'IIIIIII'III'III'Il
0 ‘00 120 ’40 ‘50 130 200 220
C1-FF0KNOWN 14 Blue FF-KNOWN

260 280 300 am 340

 

TVIrv—Y—rrr‘T—fYr'I'YTrTrrYYTIYr'1TV
0 100 120 I40 160 180 260 280 300 320 340

800
600
400
200

 

     
 

C1-F F-KNOWN 14 Yellow FF~ KNOWN

 

(A
m E
U

C
Semi?

C1-FF-KNOWN 14 Rod FF-KNOWN

 

8

[SEE RE HE W
E :11

gm 647

300
200
100

900
600
300

 

 

 

 

82

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

ABIA_ Thesis data ~ Group 2
PRISNI Licensed to Michigan State Police, DNA Unit Genotypom 2,,
Y’T’Y‘VrllirllIYTTIYYTTrT'I'TIIVTr I'— I‘lrerIlIITrth
0 100 120 140 160 100 200 220 240 260 230 300 320 340
IIII'III'III'III'IIIIIII'IIIIIII'IIIIIII'III'IIIlIIl‘II
o 100 120 140 150 100 200 220 240 250 200 300 320 340
D4-J-KNOWN 21 Blue J-KNOWN
1000
500
El
1457
WKNOWN 21 Green J-KNOWN
2000
I | I 1000
E: l¢fifihll4fifi£ lfiiil
m m m m
WKNOWN 21 Yellow .Hotown
L000
L400
L200
[I]!!! m
777
El!
0+040KMNN 21Ru1 .+KNOM01
900
00
300

 

 

 

 

83

14.3% Thesis date - Group 3
PR Licensed to Michigan 51019 Police. DNA 0011 GenotypetO 2.1

rr-rI—rrrrrrrf‘r r rfrrT—v—r‘F—T—r’rrrrr‘rtfirr'Y—r‘rrrr'rr F1"; r7 r7; r‘r‘rrr‘

’80 100 120 140 160 180 200 220 240 260 280 300 320 340

 

11101.1111111111111101:105111111111111111;1vlgsrrgrr151
80 ‘00 120 140 160 180 200 220 240 260 280 300 820 340
CB-M-KNOWN 24 Blue Nil-KNOWN

 

 

   

 

 

 

 

 

 

 

0000
2000
1000
1
IE
DE-M-KNOWN 24 Green 1.1-mama
L4000
Laooo
52000
A 1 {31000
II] 10 SE
Da-M-KNOWN 24 Yellow wmwn .
E000
‘faoo
4r300
at!
1079
m i
050
owmmoww 24 Bed M-KNOWN
> 00
. 300
300

J ‘

 

 

84

ASIA.
PRlSM

Thesis data - Group 2

Licensed to Michigan State Police. DNA Unit

Genotypem 2. t

 

rT—T fl T—FTI 1' r7—I

I—III‘YVfI—rfrfifirthfirfirI’I—IV—Ir—TfiY—Yt—IY—TI—ll

 

 

 

 

 

 

 

 

 

ii iii Jul

00 100 120 140 150 100 200 220 240 260 200 300 320 340
I‘III’IIIIIII’IIIIIII'III’III|III|III‘IlI‘III'III'IIIII
so 100 120 140 160 100 200 220 240 260 200 300 320 340
010-O-KNOWN 25 Blue O-KNOWN
900
00
300
II; In 211
BE an
IE £15
£213 EB
DtO-O-KNOWN 25 Green O-KNOWN
1500
1000
E E] :0 E85]! {BIB
Ell EB]
BEE
DtO—O-KNOWN 25Yellow o-KNOWN
.5400
300
E200
E100
__ _
‘ an GE
an [E
m:
~010-O-KNOWN 25Red O-KNOWN
000
000
400
200

 

85

 

ABIA Thesis data - group 3
PRISM

Licensed to Michigan State Police. DNA Unit GenotyperO 2.1
I'IT—r—T [firrr‘trtlvrrerrII l'rrTI‘IIUm'rrWII'I‘TTYT

100 120 140 160 180 200 220 240 260 280 300 320

 

IIIIIIII’III'III’IIIIIII'III'DII'III'III'III‘IIIIIII
100 120 140 150 180 200 220 240 260 280 300 320
OG-U-KNOWN 23 80.10 U-KNOWN

 

 

 

 

 

 

 

 

 

 

 

 

 

 

1000
Ill 8! E
129 970 1037
D&U-KNOWN
1500
1000
500
IE EB 31-2 m
m 1122] 770m
[[3 1062
EH!
23Yellow U-KNOWN
600
00
200
E] [I] ,9!!!
700 518
1:0

 

DG-U-KNOWN Z) Hod U-KNOWN

900
00
300

 

 

 

86

ASIA. Thesis date 1 group 3
PRISM Licensed to Michigan State Police, DNA Unit Genotypero 2.1
YTTIUY'TYYI—IYIII'I'I’YII’VI[fit—I—rrrrYtfiTIIIII’YfiI’firIIUTrTI]I

80 100 120 140 160 180 200 220 240 260 280 300 320 340

 

I ' I l I ' I I I I I I I I I I I l I I I 1 I I c l | | 1 ' I‘ l 1 1 I l I I o I 1 I e I I 1 I l l 1 I l I ' I
80 100 120 140 160 180 200 220 240 260 280 300 320 340
C1 -O-KNOWN 1 4 Blue GKNOWN

 

 

 

 

800
600
400
200
IE Ill 3}
ma EB
Ell El
Ell
CPO-KNOWN 14 Green O—KNOWN
1500
1000
500
A . _ 1 -l .
IRE rm: EH! EB
Cl-Q-KNOWN 14 Yellow O-KNOWN
400
200

 

 

(a)
0
co

CLO-KNOWN 14 Red O-KNOWN

 

 

 

 

87

AB'A Thesis data - group 3
FKQESBJI

Licensed to Michigan State Police, DNA Unit Gonotypenb 2.1
Tvrrt—TIlitrlrrt[Tiflftir‘l‘r‘l'jTirltI‘I'I'w'IiIII"fi'I—IitFlt'

80 100 120 140 160 180 200 220 240 260 280 300 320 340

 

I 'II I‘I II III I'l III III |I II‘ III lII III II‘ III III III II' II l'l
80 100 120 140 150 180 200 220 240 250 280 300 320 340
C9BB-KNOWN 19 Blue BB-KNOWN

 

 

 

 

F3000
-2ooo
AAA L 11000
I13 II] III
aflifl E3333 ‘065
26
EEII
CS-BB-KNOWN 19 Gwen BB»KNOWN
1500
1000
A 500
E33
1317

   

C9~BB-KNOWN 19 Yellow ABS-KNOWN

1000

11 500
Ill

32% IE!!! 1103

   

 

900
00
300

 

 

 

 

88

ABIA Plots - c1809 results
PRBM Licensed to Lynne Helton, Michignn State Police Genotypgro 2.,
TT'TWWI‘IYTTIT‘rUTTTT—fITWI1‘TfirT1l‘YrrhfimrTlerTVixtT YTUr‘I‘TII’II ——

80 100 120 140 160 180 200 220 240 250 280 300 320 340 360 380 400

IO'III'IIIIIII‘II. ICI‘I'I'IOI'COI'IIO‘III'III‘III‘OIO'IIIlIII‘III'S

80 100 12011$0 160 180 .200 220 240 260 280 300 320 340 360 380 400

‘5 KNOWN F 5 5"“ KNOWN F
2000

1500
1000
500

 

   

A5 KNOWN F SGreen KNOWN F

000
2000

 

E m .
am
0

A5- KNOWN F 5 Yellow KNOWN F

3 .
2000
-J 1000

 

 

 

 

 

II] III III
1523| m 1957
13
am
As- KNOWN F 5 Rod KNOWN F
900
800
300
J

 

 

 

 

 

 

89

APPENDIX C

Electropherograms generated from 5, 15,
and 30 second samples.

90

Thesis date - Group 1
Licensed to Michigan State Police. DNA Unit

931%”

 

 

 

 

 

 

 

 

 

 

 

 

   

 

 

 

 

Genotypero 2.1
'1 I I V] I1 I l T—jfijfiT—fiIT—lfitIYT I l'fiV—IrTYTT Ufi'UTT—r'ti UTIWTT
100 120 140 160 180 200 220 ‘ 240 260 280 300 320 340
IUI‘IOI’III'IIIIIII'IIIIIIIll!||1|I‘|II‘III‘II||‘\I'III
100 120 140 160 150 200 220 240 260 280 300 320 34a
012.0D-5 26 Blue DD-S
; 1500
1 1000
. l 500
IE 21
[EB [661
:23
012-00-5 zeemn DD-s
71500
-1000
II 4100'
Eli
01200-5 26de DD—S
1500
1000
1 500
II] E]
497
IE
491
1312-00-55 269“ 00-5
800
600
00
200

 

 

 

 

 

91

ASIA Thesis data - Group 1
PRISM

Licensed to Michigan State Police. DNA Unit Genotypero 2.1
U‘ijIVTYIVTFTII‘IVUY'I—FVTVIU‘TTUIlY—U’IIUU‘YUY'IITT'I'I'

100 120 140 160 180 200 220 240 260 280 300 320 340

 

II III 10' III III l.l III III .II I'l II' III .II Ill II] III 'II l'l II
100 120 ‘ 140 150 180 200 220 240 260 280 300 320 340
51-00-15 27 Blue 00-15

000
800
300

 

 

E1-DD-15 27 Green 00-15

?eoo
r800
-aoo

'V

 

    

E1 -DD-15
800

800

00

‘1 _200

A *A‘L

E1-DD-15 27 Rod 00-15

00
00
200

 

1 *— L

 

 

 

92

ASIA Thesis date - Group 1
PR

ISM Licensed to Michigan State Police, DNA Unit Genotypero 2.1

 

Tirlll’lrtT‘rrfi‘ll’UljfiYWVUUIIT'IT'IIIIIIIIIITIUri—IFI'I'IU
100 120 140 160 180 200 220 240 260 280 300 320 340

III'IIIIIII'III'III'lII'III'III.II‘I'IIIIIII'I'I'llllIII
100 120 140 160 180 200 220 240 260 280 300 320 340
5300-30 238100 00-30

 
  

 

 

 

-3oo
L200
II L100
II] Ell
HE! 135
6300-30 28 Green 00-30

 

53-09-30 23 Yellow oo-ao

 

E30060 28 Red DD-m

.}eoo
r600
7400
r200

 

 

 

 

93

 

AB'A. Thesis data - group 1
PR

15M Licensed to Michigan State Police, DNA Unit GenotyperO 2.1
TrYIUIj—r‘I—TY—r Tl F—rYTT fir—VIIIYWY T711 ITTT'Y TW‘IITUY“ frl—IYYUI I TIT
10 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

’IIIIIIIIIII’III'III'III’lll'III’IIIlIII'III'III’III,III’III|
10 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
BS-N-S “Blue N-S

 

 

 

 

 

 

 

 

 

 

 

 

400
300
200
100
86-N-5 11 Green NS
600
00
200
.0 .4
IE
Ill!
BS-N-S 11 Yellow NS
200
150
100
. 50
m
247 214 El
stN-S 11Red N-5
900
600
300

 

 

 

 

94

Licensed to Michigan State Police, DNA Unit Genotypofg 2.1
f']'VVI'VVTVTYTIUfIYII'IIIIII—fIYYrIIIfifitfirriv[ItrlttrIrIIerf‘llv

BO 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

APBEAM Thesis data - group 1

 

rli'UIIIOII|OIIlllll!|0|llll""I9I'III'IIOIIIIllIIIIII'OIIIIII'II

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
BB—N-15 12 Blue N-15

20

EB
88-N-15 12 Green N—15

30
20
1 0

88-14-15 12 Yellow N-15

20
10

12 Rod N-15

T 15L U1 J :22

 

 

 

 

95

 

ABIA Thesis date - group 1
PR

18M Licensed to Michigan State Police. DNA Unit Genotyperc 2.1

 

T

TIT—lIjrrijfi‘f‘rrTT—T'ITT'r—VTfTIITjfiT—‘TFfiIY—IIri’f'lTfi’l’mYYUI—ITTII—I
80 100 120 140 160 160 200 220 240 260 280 300 320 340 360 380

tIIIlIIIlIII|IIIIIIIIIIIIIII‘IIIl1II|III|IIIIIII‘III|III'III'III'II
80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
B10-N-30 13 Blue N30

80
60

20

 

B10—N-30
90
60
30

     

810-0140 13 Yellow N-SO

20

Ill

B10—N-30 13 Rod N-SO

M L i 2333

 

 

 

 

96

AB'A Thesis data ~ Group 1
PRW Licensed to Michigan State Police. DNA Unit Genotypem 2,1
YT—rIIIIIY—T FTYITTTfiT—Y'TTjilUlTfiTj rt'rirlfli‘riiirrt‘l

100 120 140 160 180 200 220 240 260 280 300 320 340

 

IIO.III'III’IIIIIII'III'II'IIIII'III'IIIIIII'IIIIIIIIIII
100 120 140 160 180 200 220 240 260 280 300 320 340
E11-X-5 32 Blue X~5

 

 

E1 1-X-5 32 Gmn X-5

300
200
100

    
 

354

 

 

E11-X—5 32 Yellow X-5

 

E11-X-5

900
00
300

 

 

 

 

 

97

 

gagsfi Thesis date - Group 1

. Licensed to Michigan State Police. DNA Unit GenotyperO 2.1
"I'YT'IYWII IjfiT—TITfiTfYr'rI—IIW—I’IIT‘FII Vivi—11W I? I'VI‘I

100 120 140 180 180 200 220 240 260 280 300 320 340

 

IIIIIII‘III'IIIIIII'III'III|III‘III|III‘IIIIIIIIIIIIIII
100 120 140 150 180 200 220 240 260 280 300 320 340
F2-X—15 33 3100 X45

200
150
100
50

 

 

a
10 18
EB 171
F2-X-1S 33 Green X-15
200
150
100

 

 

 

 

 

 

 

F2-X-15 33 Rod X-15

 

19500
r600
1-400
L200
L

 

 

 

 

98

AB'A. Thesis data - group 1

PRISM Licensed to Michigan State Police, DNA Unit Genotypem 2.1
Yl—TITr'fTIrVVITI'Y‘rrTUTTrT—IVrrTl’I’TTTTY—rlll‘IIUIIVTIrTT—rTiijtiIl'l

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

|II||II|III|III'lll'lll|lll|lll'IIIIIIIIIII'III'IIOIDIll|ll'|ld'.I
80 100 120 140 160 180 200 220 240 260 280 300 320 340 380 380

B12-X-30 14 Blue X-30

400
300

200

1 100

 

IE ‘
EH
1

 

812-X-30 14 Green x-eo
00
300
200
1 00

 

 

B12—X—30 14 Yellow X-GO

200
1 50
100
50

 

 

Biz-X430 14 Red x-eo

5900
3500
Leon

 

 

 

99

 

PRISM Licensed to Michigan State Police. DNA Unit Gonotypons 2.1
Ij r I W I I I r' I' 1 T r r I f—FW I T—’ 1' I ‘ f r r T ' F] V I ‘7 r r I I l 1'" U l r

100 120 140 160 180 200 220 240 260 280 300 320

 

I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ' I
100 120 140 160 180 200 220 240 260 280 300 320
C1-W-5 15 8100 W-S

100
50

 

 

 

 

   

[E [E El:
121 m m
18 IE EB
m a; m
CI-W-s 15 Green WS
150
100
50
frit“ L11. - 4-* ‘LvL‘Z‘: .2222-
28 IE
29
77
15 Yellow w-s
60
o
20

.51 IE E15

Ci-W-S 15 Red W-5

J- iii 1 i 353

 

 

 

100

 

 

ABIA_ Thesis data - Group 1
PRBM Licensed to Michigan State Police, DNA Unit Genotypero 2.1
FY'IIV—IIY—FIYYIIIV—UTT—YIIIIIIY—FUIVIII’(IlllYIT'Il‘l’IlIIIVU

100 120 140 160 180 200 220 240 260 280 300 320 340

 

plI'IIOlI..Illl|lll|lll'lll'lli|lllllIlllll'lll'IIOIIII

100 120 140 160 180 200 220 240 260 280 300 320 340

F8-W-15 36 Blue W-15

200
100

 

FEW-15 as Green W115

200
100

 

36 Yellow W-15

 

F8-W-1 5 36 Red W-1 5

l l M33

 

 

101

 

AB1A Thesis data - Group 1
PR

W Licensed to Michigan State Police. DNA Unit GenetyperO 2.1
11'1I‘tl—er'I—IWYIIVV‘IT—TTIT‘I’ITITIT“I’TYUI'I’WI’IfiT‘T—IITUII

100 120 140 160 180 200 220 240 260 280 300 320 340

 

pl.IIII'III'III'lll'IlIlllI'111'III'III'IIIIIII'IIIIIII
100 120 140 180 180 200 220 240 250 200 300 320 340
F1 0-W-30 37 Blue W-ao

1500
1000
500

 

 

2000
1500
1000
500

 

 

 

  

37 Yellow W-SO

soo
800
400
200
E] [E 11;: m
HE cm
In ‘

F10-W-30 37 Red W-SO

000
00
300

 

 

 

 

 

102

ABIA Thesis data - Group 1

PRISNT Licensedvto Michigan State Police, DNA Unit Genotypoie 2.1
r—rrfi I'T' 7T1 'Trj'? l rir11 1' YTUfir'r'VU—YTT—Y—Y—I—TT'j—rrfTrT‘UTrT—I

100 120 140 160 180 zoo 220 240 260 280 300 320 340

 

Ill'\I‘O|SI||00“I||’1I1i111'IIC'I1I|IIC|I|I,I1I'UCI'1II
100 120 140 160 180 200 220 240 260 280 300 320 340
F12-AA-5 38 Blue ANS

 

90
60
30
E1
F12-AA-5 38 Green AA-S
90
00
30
13 m m
I!!!
0
IE
F12-AA-5 38 Yellow AA-S
80
60
40
20

m

F12-AA-5 38 Red M—S

800
00
00

200

 

 

 

103

 

ASIA
PRISM

Thesis data - Group 1

Licensed to Michigan State Police. DNA Unit Genotypero 2.1

 

 

YIITTYIIIUITITYTIj'TI'T'TWF—ITITUlrjIIrYTTIIIVI'YI'I‘UTITI‘fi
100 120 140 160 180 200 220 240 260 280 300 320 340
rlilllIlllllilll‘llllIli'IlI]llllllI’III|IIIIIII|III|III
100 120 140 160 180 200 220 240 260 280 300 320 340
Gt-AA-15 39 Blue AA-15
4O
. 20
G1-AA-15 SDGreen AA‘15
100
50
121i
Y
Gi-AA-15 39Yeiiow AA-1s
40
30
20
10
G1-AA-15 399w AA-15
800
00
'00
200

 

 

 

 

 

104

ABIA Thesis data - Group 1
PRISM

Licensed to Michigan State Police. DNA Unit Genotypem 2.1
III—ii—Tl—IUY—TVT'TIIYTTUIT][VIII—fillYTrlrrUlUTfltl’UITT—TITII

100 120 140 160 180 200 220 240 260 280 300 320 340

 

I l I ' I I I i I I I 1 I I l I I I I I I l I I I I I I I I I ' I I I I I I I I I I l l I I I I l 1 I I I I l
100 120 140 160 180 200 220 240 260 280 300 320 340
GS-AA-SO 40 Blue AA-30

 

 

     

300
200
100
200
100
E0
187
E0
IE
40 Yellow AA-SO
200
150
100
50
El! 0 [E
233
Ill
EE

63-M-30 40 Rod AA-SO

000
00
300

 

 

 

 

105

ésgsfi Thesis data - Group 2

' Licensed to Michlgan State Police. DNA Unit GenotypetO 2.1
'Y'i'rY—I'lrtil'rrl'l'1"I'rfififii‘rI—rITfi'lfi"Tj‘ltiilii

0 100 120 140 160 180 200 220 240 260 280 300 320 340

 

' I I I ' I I I I I I I | I I I I I I I I I I I ' I I I I I I I I I I I | I I I I I I I I I I I | I I I l I I
O 100 120 140 160 180 200 220 240 260 280 300 320 340
BS-FF-S 11 Blue FF-S

1500
1000
500

 

t1soo
1000
--500

 

 

b

r600
r400
r200

 

 

 

BB-FFé 11 Rod FF-5

900
00
300

 

 

 

 

106

Thesis data - Group 2
Licensed to Michigan State Police, DNA Unit

ASIA
PRISM

Genotypem 2.1

 

 

I'
TTIIIlITITIITITTrj—r‘irjl’r‘l’r‘I—IY'YTr—tfi‘tY—l‘TIIWTrI‘U’I’rI

 

   

 

 

 

 

 

 

 

o 100 120 140 150 150 200 220 240 260 280 300 320 340
blIl'III'III'III'III'III‘III'IIIIIIIIIIIIIIIIIII'IIIIII
o 100 120 140 160 180 200 220 240 260 230 300 320 340
Bto—FF-ls 12 Blue FF-lS
800
600
on
200
@707“
Bio-FF-is 126mm FF-IS
aoo
.,eoo
.400
9200
E in IE
EE ES
888 EB 423
B10-FF-15 12 Yellow FF—15
300
200
100
m m
EB IE
B10-FF-15 12 Red FF—15
900
600
300

 

 

 

 

107

ABIA- Thesis data - Group 2

PRISM Licensed to Michigan State Police. DNA Unit Genotypero 2.1
TUIIUIYIY—rT‘I—r'III—I—TIII'rr"Irt'rl’"ljrrjt—‘rrTV—I‘ITT'T‘r

o 100 120 140 160 180 200 220 240 260 230 300 320 340

 

'IIIIIII'III.IIIIIIIIIII'III'III'IIIIII‘IllII'III'IIIIII

0 100 120 140 160 180 200 220 240 260 280 300 320 340
B12-FF-30 13 Blue FF-ao

300
200

100

       

IE to
En

B12—FFo30 13 Green FF-SO

L400
L300
L200
3100

 

 

 

13 Yellow FF—GO

90
60

80

EDIE

81 2-FF-30 13 Red FF-ao

800
00
00

200

   

 

 

 

 

108

ABIA. Thesis data - Group 2

PRISM Licensed to Michigan State Police, DNA Unit Genotypeno 2.1
'T' ‘I—Yj—Y'V'ITTTI' T—‘T'T' T I I t T Y I VT. rlfirrt I I Y Y'IT' t ' I I ‘I—fI I '—'

100 120 140 160 180 200 220 240 250 230 300 320 340

 

I I I ' I I I ' I I I | I I C I I I I ' I I I I I I . I ' . I i I I I | ' I I ‘ | I I ‘ ‘ I I l I I . I I I I
100 120 140 160 180 200 220 240 260 280 300 320 340
CQ‘J-5 13 Elm J-S

150
100
50

 

 

69-4-5 18 Yellow J-5

 

094-5 10 Rod J-5

9000
9000
5300

b-

 

 

 

 

 

109

ABIA_ Thesis data - Group 2
PRISM Licensed to Michigan State Police. DNA Unit acnotypcro 2.1
7W,r'I'T—‘T't'r'TTYT—YTVTYr'r"I'Vr‘Trfi—YT" VIIII‘leIrTUU

100 120 140 160 no 200 220 240 260 230 300 32o 340

 

II!‘III'III‘III‘III'III'IIIIIIl‘lll'lll'llI‘IOI'III'III

100 120 140 160 180 200 220 240 260 280 300 320 340
GHQ-15 19 Blue J45

 

 

 

 

 

 

 

 

 

1500

1000

500

k L k _ +
m
m
[E
m
C11+15 19W J45

2000

1500

P‘OOO

. A -500

IE
m
795
C11415 19 YOIIOW $15

00

00

200

m
745

 

0114-15 19 Rod J-15

in: J J 35‘?

 

 

 

110

 

ABIA Thesis data - Group 2
PRISM

Licensed to Michigan State P0lico.-DNA Unit Genotyporo 2.1
lIler'rrtlIITr'rfij’r—YII'UI‘VY'I'I’V‘UIUI’IIYVU'UY'Ir‘TrI'I

100 120 140 160 130 200 220 240 260 280 300 320 340

 

100 120 140 100 130 200 220 240 250 280 300 320 340
024-30 ZOBluo J-ao
1500'

1000

I I 500
Iii
EEIJ
II]

 

 

 

024-30 20 Gwen +30
1500

1000
00

 

 

490

txa+eo zowwflmu.400

#000
L400
E200

 

 

 

024-30 20 Rod J-m
800
00
00
200

 

 

 

 

111

ABIA, m. m . em 2
PRISM Licensed to Michigan State Poison. DNA Unit Gonotyporo 2.1
rIVTItIIrrT'IIVrTYYTIYTTI'T’TY—V'TTYtY—IYI’TIUW"TTI'I'IITI

0 100 120 140 160 180 200 220 240 260 280 300 320 34'!)

 

IIIO|III'IIIIIIIIIIO'III'Ullllll'lll'lll'lllIIIO'IIOIII

0 100 120 140 160 180 200 220 240 260 280 300 320 340

AT-M-s SBiuo NM)
30
20
10

.H l ‘ , , i ‘ A l I . I '

A7-M-5 5 Gmn M-S

20

AHA-S 5 Yellow 01.5
30

20
10

A7-M-5 5 Rod M-5

 

 

 

 

112

AB'A Thesis data ~ group 2
PRISM Licensed to Michigan State Police. DNA Unit GanotyparO 2.1
YrYIY'TfI'I{Uri—T'VfirYrr'UrVTVTTr'YVIYIT—II’TIIIIVIIYTIYTII'IVII

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

PIIIIIIIIIIIIIIIIIIIIIl.l0|'lIlIltllllt'lll|IOIIlll'llIllllll
100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
A7-M-15 5 Blue M45

 

   

 

 

 

 

 

 

 

 

400
300
200
‘1 L ‘°°
’
EB
IE
HE
A7-M-15 SGmon M-15
}000
7800
740°
200
A7-M-15 s Youow M-15
00
0
30
m n;
[E13 115
IE Ill
[IE EB
A7-M-15 5 Rod M-15
900
600
300

 

 

 

113

 

Ucensed to Michigan State Police. DNA Unit GonotyperO 2.1
fY‘YVTIIIIrTrIIITTI‘YrT—rTUIliiflVrI‘TIUITI—TITT‘r' '1'“. UI'VUTI'

$00 120 140 160 180 200 220 240 260 200 300 320 340 350 380

Ang%- Thesis dais - group 2

 

I.IIIOI.OlllIII.OII'III'III'lll'lllllll'lllllll'IOI|III|IOIII
100 120 140 150 180 200 220 240 260 280 300 320 340 350 380
AQ-M‘m BBIUO M-30

O
80
40
20

II}

E?

IE
ADM-30 8 Gwen M-ao

 

900
600
300

 

 

 

 

114

ABIA Thesis w. . Group 2
PRW Licensed to Michigan State Police. DNA Unit Genotypeio 2.1
fivtltT—‘I—IYVTIYIT]Ir111rfritrIITf'IVTIYY‘TIfIVfiYIIIYY'rIV

0 100 120 140 160 160 200 220 240 260 280 300 320 340

 

llll'lilIIII'lll'lll'll‘l'il'llllI'I'lillllll'llfi'lillll
O 100 120 140 160 180 200 220 240 260 280 300 320 340
82-0-5 88iue 0-5

30
20

82-0-5 8 Green 05

20

82-0-5 8 Yeliow 0-5
30
20
1 O

32-0-5 8906 0-5

900
00

 

 

 

 

115

ABIA_ Thesis data - Group 2
PRISM Licensed to Michigan State Police. DNA Unit Genotypem 2.1
TIIII'IT—rrT—ITIrIITIIIIIIITI'IIIIYYIIVIIrT—YIITTV['Irrtr

O 100 120 140 160 180 200 220 240 260 280 300 320 340

 

I I I I I I I I | I I I I I I I I l I I l I I I ' I I I I I I I I I I I I I I I ' I I I I I I I I I I I | I I
O 100 120 140 160 180 200 220 240 260 280 300 320 340
34-0-15 9 Blue (>15

40
.2 20
a! :3

84-0-15 QGreen 0-15
0
0
0
20

EB E

84-0-15 9Yeiiow 0-15
0
20

:2

84-0-15 980d 0-15
800
600
400
200

 

 

 

116

Thesis data - group 2
Licensed to Michigan State Police. DNA Unit GenotyperO 2.1

TU'IIYITIIIIUVI‘IITTIIYIYVIIIII‘IrT'fiITTIIII]TUW'IIII'1'

100 120 140 160 180 200 220 240 260 280 300 320 340

ABIA
PRISM

 

100 120 140 160 180 200 220 240 260 280 300 320 340
560-30 10 Blue 0-30

 

0
20
IE
m
380-30 10 Red 030
000
00
300

 

 

 

 

 

117

ABIA Thesis data ~ Group a
PRW Licensed to Michigan State Police, DNA Unit Genotypero 2.1
I‘YIYI‘III'IIrII’rIII’IIIrl‘IIITTT—YIrIYIrV'I—vTrVYIIII'UII

100 120 140 160 180 200 220 240 260 280 300 320

 

III.III|III|III|III'III[III'III'III'III'III'III'III

100 120 140 160 180 200 220 240 260 280 300 320

Ct t-U-5 20 Blue U-5
2 0

15

C11-U-5 20 Green ch
4o

30
20
1 0

 

   

Ct 1-U—5 20 Yellow U-S

 

C1 1-U-5 20 Red U-S

 

 

118

 

ABIA,
PRISM

Thesis data - Group 3

 

0241-15

D2-U-15

   

Licensed to Michigan State Police, DNA Unit Genotypeto 2.1
I'IITII‘VIIY'T'IUI'UT'I'Tr'l'tlTII'IUrTII—UIIiI'IIr
100 120 140 160 180 200 220 240 260 280 300 320

III'III'III'III'III'III’III'III'III'III'III'III'III
100 120 140 160 180 200 220 240 260 280 300 320
D2-U-15 21 Blue U-15
BO
60
0
20
m E! E m
'02-U-15 21 Gwen U-15

 

21 Yellow U-15
40
20

m

21 Red U-15

_U L A y SEE

 

 

 

 

 

119

ASIA. Thesis date - Group 3
PRISM Licensed to Michigan State Police. DNA Unit Genotypero 2.1
'UIj—TfitiTFIITIIVrTjtTI—rVIIIUIIIIIIrV‘Ufi‘ITT'Tir'Yrr

100 120 140 160 180 200 220 240 260 280 300 320

 

I I I I I I I I I I I | I I I ' I I I ' I I I I I I I I I I I I l I I ' I I I I I I I I I I I | I I I
100 120 140 160 180 200 220 240 260 280 300 320
04-an0 228iue U430

0
‘. 20
‘ i , .' i
51-: Hi El
04-an0 226mn U-30
80
0
0
20
B [E
El!
[E
El
D4~U~3o 22 Yellow U-ao
0
30
20
10

22 Red U-3O

04-11-30
900
00
300

 

 

 

 

120

APEIA Thesis data - Group 3

ISM Licensed to Michigan State Police. DNA Unit GenotyperO 2.1

 

fi'rvtrTIfTTrfi‘erTrl’ITI—YwI—TI—‘IIIfi’IYrTIITT—rtjfjiriIr
100 120 140 160 180 200 220 240 260 280 300 320

III'III'III'III'III'IIIIIIIIIII'III'III'IIIIIIIIIII
100 120 140 160 180 200 220 240 260 280 300 320
88-0-5 118iue 0-5

mm m
32
M

   

 

 

88—0-5 11 Green 0-5
IE [E] [D
me an a;
88—0-5 11 Yellow 0-5

m a m (I:
m
IE m
ma w
“Red 05

“i 1111 J

 

200
100

 

100
50

 

 

 

121

 

ABIA_ Thesis data-Group 3

PRISM Licensed to Michigan State Police. DNA Unit Genotypes-O 2.1
rr'r'ItTl[IYTIF'rr'T—T‘TYfiTITIIrl—IilrIrTj‘II'llIII’VU'

1 DO 120 140 160 180 200 220 240 260 280 300 320

 

III'III'IIIIIIIIIIIIIII'III'III'IIIIIIIIIII'IIIIIII
100 120 140 160 180 200 220 240 260 280 300 320
810-045 12 Blue 0-15

      

 

 

 

 

150
100
so
IIEIIEEI IIEIIIi
[IE] III!
II!
810-045 12Green 0-15
00
300
200
1 100
ill IE 3 IE m
Hill :53 144 £21 I!!!
Ell
III]
810-0-15 12 Yeliow 0-15
150
100
50
Ill .13 I12
Ilia III -li§
Ila
Ill
910-015 12 Red 0-15

 

P900
. }600
E300

 

 

L22

 

ABIA Thesis data . group 3
PR

ISM Licensed to Michigan suns Police, DNA Unit GenotyperO u
1'1Itt'ftl[III]IT!'IITTrUVITijUIYTW—UTII—T—TTTFU'III

100 120 140 160 180 200 220 240 260 280 300 320

 

I I I I I I I | I I I II I I I I I I ' I I I i I I III I I I I I I l I I I | I I I | I I I I I I I
100 120 140 130 180 200 220 240 260 280 300 320
812-030 138iue 0-30

}400
~300

ll 9200
7100
32
E!!!
E15
E15]

 

 

 

 

 

 

 

 

 

 

£113
B12-O-m 13 Green 0-30
E‘ m
Elli 1113 £123
Ell
EEK}
812-030 13 Yellow 030
. 300
200
_#uh“ 100
[I] [3 I15
EIEJ E2!) 171
IE}
EIEJ
8120-30 13 Red 030

 

. P900
' }s00
5300

 

 

 

 

123

ASIA Thesis data - group 3
PRISM Licensed to Michigan State Police, DNA Unit Genotypene 2.1
I 1 I T 1’1 r I T I I I "l 0 I I I ‘l' I” T l U I T 1 Y 1' ‘ I T f I I r I I W r Y I I ' ‘ ‘ I I j

100 120 140 160 180 200 220 240 260 280 300 320

 

IIIIIII'IIIIIII'III'Ilt'lllllII'IIIIIIIIIIOlIIl'III
100 120 140 160 180 200 220 240 260 280 300 320
(Xyaafi HIBWO 336

 

.L300
1200
-1oo
I13 II] E!)
1358 I!!! III
E13
1!
03-885 16 Green 88—5
200
150
100

 

 

(XÂ¥886 16th! 8845

~1000
#500

 

 

 

 

12Ai

 

ArggsAM Thesis data - group 3

. Licensed to Michigan State Police. DNA Unit Genotypem 2.1
ll'I‘l"[Uif‘lfilvifiITIIIY'I’TIYY—[T'VthIIrYTr!IIITYT

100 120 140 160 180 200 220 240 280 280 300 320

 

lll'lll'lllIIII‘III'OII'IIC'III.IOI'III'lll'llllIt!
100 120 140 160 180 200 220 240 260 230 300 320
05-8845 17 3'00 35-15

 

 

L400
L200
J__ _ __ __ ‘U .
m *
517 a [m
E
Hi

0586-15 17 Green 88-15

 

 

 

 

05-88-15 17 Rod 88-15

900
00
00

 

 

 

 

125

ABIA Thesis data - Group 3
PRISM Licensed to Michigan State Ponce, DNA Unit Genotypcie 2,1
'7 I I I I I r I r I r I I I r 1 I I I I I I U I I I l I I I I I I I I I I I I I I j I I I I I I I I

100 120 140 160 180 200 220 240 280 280 300 320

 

III'III'III'lIO'IUI|III'IIIIIDD'DII'OIO'IIIIOIIIIIO
100 120 140 160 180 200 220 240 260 280 300 320
(”-38.30 18 Blue 88-30

 

 

 

 

 

 

 

 

 

 

 

oo
00
200
I. A JLLJI ._ - ., c__
IE ID ED
736 716 [[33
[EB
07-88-30 18 Green 38-30
00
r400
haoo
a i
559
Em
(27-88-33
200
I 100
IE
[IE
07-88-30 18 Red 83-30

 

- Leon
E-eoo
gaoo

 

 

 

 

126

ABIA
PRISM

Thesis data - Group 2

 

 

 

 

 

 

 

 

 

 

 

Licensed to Michigan State Police. DNA Unit GenotyperO 2.1
rIr'IIIUTfIIFYITT'TT'IYI'WTI'— T'IUU'YIYIII'II'YI’III’YIIIT
100 120 140 160 180 200 220 240 260 280 300 320 340
llI'lll'lllIlll'lll'lltlllI'llI'III'IIO'OIIIIII'III|Ill
100 120 140 160 180 200 220 240 260 280 300 320 340
DS—F-S 23 Blue F-S
300
200
I I I 100
IE 18 E
114 “E
[E 2::
EH] [IE rm
DS—F-S zaGreen F-5
Leoo
H00
I 200
J A; M L 4
E III [E]
739 E
IE
06-F-5 23Yeiiow F6
150
190
50
800
00
00
200

 

 

127

 

AB'A Thesis data - Group 2
PRW Licensed to Michigan State Police, DNA Unit GenotypeiO 2.1

IVIUTITYVYIIiinII’T‘I—TIIIIIII'ITTTYITI—‘Vr—II'T'IIY'IIT'III

100 120 140 160 180 200 220 240 260 280 800 320 840

 

pll'Ill'lll'llIlIll'lll'llI'Oll'lllllll'III'IIIIIIIIIOI
'00 120 ‘40 150 180 200 220 2‘0 250 280 300 320 340
03-F'15 24 Blue F-15

MOO

L
l I ~200

 

 

 

 

 

 

 

 

 

 

 

 

Eli
EM
034-15 24 Gwen F15
19000
igeoo
I I 9300
E in EE
an em
03-315 24 Yellow F-ts
' zoo
200
I 100
m
[EB ma EB!
IE
rm
oe-Hs 24 Red F-15

 

.Esoo
r800
.400
5200

 

 

 

 

128

Thesis data - Group 2
Licensed to Michigan State Police, DNA Unit

ABIA
PRISM

Genotyper® 2.1

 

F0 100 120 140 160 180 200 220 240 260 280 300

1 6 0
F-SO

100 120 140

9 Blue

30 180 200 220 240 260 280 300

B44230

 

 

 

B4-F-3O 9 Green F-30

In!
M

 

34-53-30 9 Yeliow F-30

 

'Ill'illllll'llll'llllllllll'lll'lll'lllII'O'III'

320 340

320 340

.- A“ _.__

‘rK—Il'Vfi—TTTUVTTTVIFIVvaiiljIle‘YTTV'IIrrTIIIITjV‘UIlfl‘IVUT—I

360 380

360

380

300
200
100

600
00
200

~200
~150
~100
r-SO

 

 

—— v

 

 

344-30

800
600
400
200

 

 

 

129

 

AB'A Thesis data - Group 2
PRISM Licensed to Michigan State Police. DNA Unit Genotype“ 2.1
I'IIVTIYY ‘IIU—[fi—YTYUIli'r'rlr1W117TYTWrV'W YT'rTYIYIVIITT'

l0 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

'III'III’III'llI’llI'lll‘llI'Ill'lll'lllllll'lll'lll'III'III'
0 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
BZ-L-S 8 Blue L-5

 

 

BZ-L-S 8 Green L-5
200
100
x a
262 m E E m
£19
an
BZ—L-s a Yellow L-5
0
40
20
EE
Bz-L-S
900
00
300

 

 

 

 

130

ASIA- Thesis date . Group 2
PRISM Licensed to Michigan State Police, DNA Unit GenotypeiQ 2.1
I' ' T [fir T r IfT T l I I t Ij—T’fi I I I 1' hr 'j 1' I ' lfi 1 Ij I ' T—t I' ‘7 r F‘ V r r I f‘ I 1'

0 100 120 140 160 180 200 220 240 260 280 300 320 340

 

' I I I l I I I ' I I I I I I I I I I I I I I I l I I I ' I I I I I I I I I I I I I I I I I I I | I I I I I I
O 100 120 140 160 180 200 220 240 260 280 300 320 340
CS-L-lS 16 Blue L-tS

00
200

 

8%
El

595 iii]
“:1

 

a
‘1
D

CS-L-ts 16 Green L-15
.300

E-BOO
MOO

r200

 

 

   

C5-L-15 16 Yellow [:15

 

C5-L-15 18 Red L45

9900
.—500
-aoo

 

 

 

 

 

131

ABIA Thesis data - Group 2
PR

ISM Licensed to Michigan State Police, DNA Unit GenotyperO 2.1
VITIIUIITT'Y—f I IrrIIIIIVY‘fiIY'rIIIIIIITWITI’TT' TUT'fIfi'I
O 100 120 140 160 180 200 220 240 260 280 300 320 340

 

p I I I | I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
0 100 120 I40 160 180 200 220 240 260 280 300 320 340
C7-L-30 17 Blue L-ao

 

 

 

 

 

 

 

 

 

   

800
600
00
Ml . I I 200
+ A) AA AA A AAAAA A AA A 7
IE 18 m
«0 an
IE m
em
C7-L-30 17 Green L-30
1000
500
A. L A
12
144
13
187
200

. I 100

 

 

 

95
IE
[EB
C7-L-30
900
000
300

 

 

 

 

 

132

ABIA
PRISM

Thesis date - Group 3

 

 

 

 

 

Licensed to Michigan State Police. DNA Unit Genotype“ 2.1
IIIUT'III'IIT'TYIT'IrIrj'IIIYlT'IlIT,IIIW'TjI'I'I
100 120 1 40 160 180 200 220 240 260 280 300 320
IIIIIII'III'IIIIIII'IIIIIII'IIIIIIIIIIIIIIIIIII'III
100 120 1‘0 160 180 200 220 240 260 280 300 320
8244-5 BBiue HS
150
100
50
8241-5
300
200
l 100
III?
IE
[IE
aa-u-s 8Yellow 1+5
80
0
0
20
8244-5
900
00
300

 

 

 

 

 

 

133

ASIA Thesis data - Group 3
PRISNT Licensed to Michigan State Police, DNA Unit Genotypero 2.1
Tl'fV'ITrII'Tt'W‘IrrTViI’I'IU'fIfiIIIrTrUIITYII'I’II’Y'

100 120 140 160 180 200 220 240 260 280 300 320

 

I II I' II I' II II I a. II a. II n .0 II II II II II II II I‘ I u. II r. II I
100 120 I40 150 180 200 220 240 250 280 300 320
M15 9 Blu. H45

800
800
400
200

 

 

-1000
$500

 

 

i2

   

M15 9 Yellow H45

‘r400
~300
#200

r100

 

 

8441-15 ,

 

 

 

 

 

134

 

ASIA
PRISM

Thesis data - Group 3
Licensed to Michigan State Police. DNA Unit

Genotypero 2.1

 

86-H-30

864-1-30

10 Blue H~3O

 

‘10 Yellow H-30

10 Red H-30

 

90
60
30

100
$0

80

4O
20

900
600
300

 

 

 

 

 

135

 

APPENDIX D

Electropherograms generated from samples
containing 3ul of PCR product.

136

ABIA Thesis data - Group 1
PRISM Licensed to Michigan State Police, DNA Unit Genotyperfl 2.1
rrIIIrTrTI—TIIITTITTIII—rrrI‘f‘I'IfTIITTTITrhrr'YT1"I’rllI

60 80 100 120 140 160 180 200 220 240 260 280 300 320

 

IIIIIIIIIIIIIIIIIIIIIIIIIIIIII'III'III'IIIIIIIIIIIIIIIII
60 80 100 120 140 160 180 200 220 240 250 280 300 320
8344.15 5 BIUO N-15

200
100
IE IE
88-N-15 5 Green N-15
150
I00
50
EB 31
ea-N-is 5 Yellow N-15 _
150
100
50
III III
EB
BB-Nds 5 Red N-lS
600

00
200

 

 

 

 

137

 

ABIA Thesis data - changed amp. volume
PRISM

. Licensed to Michigan State Police. DNA Unit Genotypero 2.1
VIII!IIIITTII[IIrIIIITTrTITI'YIIT—T'YII'IITrfiI[IIII

100 120 140 160 180 200 220 240 260 280 300 320 340

 

I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I l
100 120 140 160 180 200 220 240 260 250 300 320 340
A5-X-5(3UL) 4 Blue X—5(3UL)

00
200

‘J‘A ‘ A

 

A5-X-5(3UL) 4 Green X-5(3UL)

 

 

 

 

 

 

 

 

 

 

IE [E E
699 Km HE
30.2
cm.
A5-X-5(3UL) 4 Yellow x-5(3u1.)
200
150
100
1 o
m. E
El! HE
[E In
IE3 IE
A5-X-5(3UL) 4 Red X~5(3UL)

00
300
200
100

 

 

 

 

138

ABIA. Thesis date - changed amp. volume
PRISM Licensed to Michigan State Police, DNA Unit GenotyperO 2.1
jji—YT—rY—rTITIt—rtliifillffifiIfI’IIIrIYI'TI'T‘ftlUfiTIII’IT

80 100 120 140 160 180 200 220 240 260 280 300 320 3‘0

 

I I I I I I I I I I ' I I I I I I I I I l I I I I I I I I I I I I I | I I I ' I I I '~I I I I I I I I I I I I
so 100 120 140 160 180 200 220 240 260 260 300 320 34c
A7—X-15(3UL) 5 Blue x-zswuu

00
300
200
100

 

 

 

A7-X-15(3UL) 5 Queen X—15(3UL)

‘9400
IrSOO

200
E100

 

 

 

 

A7-x-15(3UL) 5 Yellow X—15(3UL)

 

#‘ ‘ LAL ___ AA A

   

A7-X-15(3UL) 5 Rod X-15(SUL)

E400
~3oo
L200
P100
L-

 

“‘ ‘A‘.‘
A L ‘- A

 

 

139

 

9R§ISIAW Thesis data - changed amp. volume

 

Licensed to Michigan State Police. DNA Unit Genotypeto 2.1
‘I I l I I I I I T rT T’j’l I—r f I l I I l I 1" I Y T '1 fr I I I I I I I I T I T T r I
100 120 140 160 180 200 220 240 260 280 300
I I I I I | I I I . I I I ' I I I | I I I ' I I I ' I I I I I I I | I I I I I I I | I I I
100 120 140 160 180 200 220 240 260 280 300

BSoW-15(3UL) 12 Blue W-15(3UL)
900

l 00

“I 00

W 788
IE
an

A;

 

 

 

 

m
18

ea-w-wwuu 12 Green w-vswuu

Egon
on
400

L {-200
IE

 

 

Ba-W-15(3UL) 12 Yellow W-15(3UL)

A no
‘ zoo

“2
an 451 EB £33
In

sew-mam.) 12 Red W-15(3UL)

 

 

 

 

{-400
r300

200
L“00

 

 

 

140

 

ABIA Thesis data - Group 1 (re-run)
PRISM Licensed to Lynne Helton, Michigan State Police Genotypero 2.1

rIIY—‘rTjrlIrTTT'IIIjr‘FTII’TIjjIII—IITI‘ITT'IIIII
100 120 140 160 180 200 220 240 260 280 300 320

 

IIIIIIlIIIIIII'III'III'III|IIIIIIIIIIIIIII'IIIII

100 120 140 160 180 200 220 240 260 280 300 320

DG—AA-S 16 Blue AA—S

 

 

 

   

 

 

 

.200
100
A AA- - L LT;
DB-AA-S 16 Gwen AA-S
200
150
I00
50
E IQ m
E *3 IE
I] IE E
[E E1! [IE
DO—AA-S 16 Yellow M-S
200
150
100
n - - g; A» LJL- 5°
III E III IE
237 me E!

DB-AA-S 16 Rod AA-S

on
200

 

 

 

141

 

7 ABIA Thesis dale - changed amp. volume
PRISM Licensed lo Michigan State Police, DNA Unit GenotypetO 2.1
U V f T I 1' I r I I I ‘ T t I I I I 1 r r—' I l I I 'j I I 1 I 1 r I I j IV r [T ‘ Ir—i

100 120 140 160 180 200 220 240 260 280 300

 

o | o I o | v I I | n I I I I 1 I ' 1 I I I I 1 I I I I 1 1 o I n I I I 1 I o 1 a ' 1 1 1
100 120 140 150 180 200 220 2‘0 260 280 300
BB-M-30(3UL) 11 ENG AA-30(3UL)

200
150
100
50

gagla

 

Bis-“mum 11 Gwen AA-30(3UL)

150
100
50

   

150
100

 

BB-AA—30(30L) 11 Red M-30(3UL)

00
300
200
100

 

 

 

142

 

ASIA Thesis data - changed amp. volume.
PRW Licensed to Michigan State Police. DNA Unit Genotypeco 2.1
I I I r I l I I 1 T T I I I I I I I—I' f I I I I I I I T 1 I I I I l' I I T I I I—j' r I I I

100 120 140 180 180 200 220 240 260 280 300

 

 

 

 

 

100 120 140 160 180 200 220 240 260 280 300
B12-FF-30(3UL) 14 Blue FF-30(3UL)
00
00
200
RE 513 5E1 EB
B12-FF-30(3UL) 14 Green FF-30(3UL)
00
00
00
200
um
B12-FF~30(3UL) 14 Yellow FF-30(3UL)
200
150
100
50

Elm

B12-FF-30(3UL) 14 Red FF-30(3UL)

7400
7300
7200
r100

 

 

 

 

 

143

 

ASIA.
PRISM

Thesis data - changed amp. volume

 

   

ANSBU L)

ANSQUL)

w
HE 3E

 

 

a Yellow J-5(3UL)

mm
Im-

a Red J-5(SUL)

@BQB

 

 

Licensed to Michigan State Police, DNA Unit Genotypeifi 2.1
TIfiIIIII‘T'TT‘I’IIII’jfiI’I—l—I—IIITITIIffir'fi'jIIII T—‘II'
100 120 140 160 180 200 220 240 260 280 300
I'IIIIIII'III'III'III'III'IIIIIII'IIIIIIIIIII
100 120 140 160 180 200 220 240 260 280 300
A9-J-5(3UL) 6 Blue J-5(3UL)
300
200
100 ‘
ll! El
257 E12
IE IE
Eil 3E
A9-J-5(3UL) 6 Green J-5(3UL)

00
400
200

150
100

?‘°°
>300
fizoo
E100

 

 

144

 

ABIA thesis data . changed amp. voiume
PRISM Licensed to Michigan State Police. DNA Unit Genotypem 2.1
IIIIrffilTTTIrIIIIIIITTIIITYIU'I—TIIIl—TITT‘IIII'T"'rlI—rIT

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

I I I I I I I I I I ' I I I I I I I ' I I I l I I I I I I I ' I I I I I I I ' I I I I I I I I I I I I I I I I l I
100 120 140 150 130 zoo 220 240 260 280 300 320 340 360
Ci-M-5(3UL) 15 Biue M-5(3UL)

 

60
4O
20
C1-M-S(3UL) 15 Green M.5(3UL)
1 00
50
E El 30
C1-M—5(3UL) 15 Yeliow M-5(3UL)
' 0
20
CI-M-5(3UL) 15 Rad M-5(3UL)
400
200

 

 

 

145

ABIA- Thesis data - changed amp. volume ,
PRISM Licensed to Michigan State Police. DNA Unit Genotypam 2.1
r I I ' t I l7 I I I I f I T I T I '1 I T- I I I I I l ' I T I T t I I 1 I ' T r I I l I I t I I I U r I T I ‘ I r

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

I I ' I I I ' I I I | I I I ' I I I ' I I I l I I I ' I I I I I I I I I I I | I I I | I I I ' I I I i I I I I I I
100 120 140 160 180 200 220 240 260 280 300 320 340 360
CS-O-S(3UL) 16 Blue 05mm.)

40
30
. , 20
. l 1 0
' , I i. A . ‘ v
03-05(3UL) 16 Green O-5(3UL)
60
- 0
20
03-05(3UL) 16 Yeflow O-5(3UL)
20
1 0
' l _ I I i I '1
ca-o-suauu 18 Red O-5(SUL)
00
200

A ‘ A- M _ _- A -

 

 

146

 

Thesis data - changed amp. volume
Licensed to Michigan State Police. DNA Unit

ABIA
PRISM

Gonotyperfl 2.1

 

r I I Y

160

I' I j

180

[YTTIITIIIYI'I—V‘I'I

200 220 240 260 280

V I I T I I I

100 120

I I' I I I

140

an'
280

I
260

220 240

140 160 180
13 BIUO U15(3UL)

200

' I ' I
100 120
B10-U-15(3UL)

HEM [EB
[E

13 Green U-15(3UL)

B10-U-15(3UL)

 

B10—U-15(3UL) 13 Yellow U-15(3UL)

53
UI

 

B10-U-15(3UL) 1 3 Rod U-15(3U L)

 

I' ‘ I’ I' I

300

300

200
150
100
50

00
300
200
100

30

00
300
200
100

 

147

 

ABIA_
PRISM

thesis data - changed amp. volume
Licensed to Michigan State Police, DNA Unit

Gonotypom 2.1

 

 

 

 

 

 

 

100 120 140 160 180 200 220 240 260 280 300
II'OII'lII.IUU'IDI'O|I'Illtllt'lII|lll'
100 120 140 160 180 200 220 240 260 280 300
CS-U-30(3UL) 17 Blue U-30(3UL)
m
m m [:5 ER (in
CS-U-30(3UL) 17 Gwen U-30(3UL)
X E
161 m
63
cs-U.30(3UL) 17 Youow U-30(3UL)
m
cs-u-aomuu 17 Rod U—30(3UL)

320

320

340

340

II'VW'UrIfi'IF‘IIjTW'IIIIIT‘IIWIYrYYYI'UIIU'I'I'UT‘I'T'UIII

330

360

80

40
20

150
100
50

60

00
200

 

148

 

APBIA Thesis data - changed amp. volume

 

 

 

 

 

 

   

Licensed to Michigan State Police. DNA Unit GenotypetO 2.1
I I I V I I ‘l T I l I T r I Tfi‘ T_' fil 1 l l i I I T I I I r I I I T 1' Ti Y I I T—I' I I
100 120 1 40 160 1 80 200 220 240 260 280 300
I I I I I I I I I I I I I I I I I I I I I I I l I l I I I ' I I I I I .I I I I I I | I I l
100 120 140 160 180 200 220 240 260 280 300
Att-L—15(3UL) 7 Biue L-15(3UL)
900
600
L 300
@-
IIIIE Ilia
II]
E113
Att-L-15(3UL) 7 Green L-15(3UL)
~1500
~1000
1-500
A A _ LA
32
EIIE
E33
317
Att-L-15(3UL) 7 Yeliow L-15(3UL)
L300

 

 

.I E200
‘ I 7100
- m.
Ei33
I13
E113

 

A11-L-15(3UL) 7 Red L-15(3UL)

L400
L300
5200
rtoo

 

 

 

 

 

1119

 

ABIL
PRISM

Thesis data - changed amp. volume

 

 

 

 

 

 

 

 

 

 

 

Licensed to Michigan State Police, DNA Unit GenotyperO 2.1
V I ‘l l’ l I I l I 1' I I I I I 1' I I I I T I I V ‘I I I I I I I I T I I T‘I lfi' T I I 1 I’ r
100 120 140 160 t80 200 220 240 260 280 300
I'III'III'III'III IIIlIII'IIIIIIIIIII'IIIIIII
100 120 140 160 180 200 220 240 260 280 300
82-88-5(3UL) 9 Blue BB-SlSUL)
r800
L400
L200
*4 M ‘ - M L - ‘ ‘A ’
EB IE!
82-88-5(3UL) 9 Green BB-S(SUL)
300
200
100
“.11 EB IE
an E1]
IE EE IE
[E III] a:
Bz-BB-S(3UL) 9 Yellow BB-S(3UL)
200
150
100
50
2in E13
82—88-5(3UL) 9 Red BB-S(3UL)
-r400
7300
7200
100

 

 

 

150

 

Thesis data - changed amp. volume
Licensed to Michigan State Police, DNA Unit

ABIA,
PRiSM

Genotypero 2. t

 

1 ‘l I I I I I I I I I I I I I I I I r I r I

fIIIrfiIIrIIIIII'IfiI—Iert

 

 

 

 

 

 

100 120 140 150 180 200 220 240 250 230 300
I'lII'III'III’III'III lll‘lll'lIlllll'lllllll
100 120 140 160 180 200 220 240 260 280 300
mmmu to Blue 015(301.)

400
200
52
BE EB GE :25
E?
m
54415900 10 Green 015(301.)
1500
1000
500
L - in - 44L
[2 IE E13 IE [E
E!!! am 470
E]!
m:
34-0-15(30L) 10 Yellow 0.15901.)
00
200
am m
an an an:
an
elm-1590:.) to Red 0-15(3UL)
400
300,
200

100

 

 

 

 

 

151

APPENDIX E

Electropherograms generated from samples
containing 3ul of concentrated PCR product.

152

ASIA. Thesis data - concentrated amp. product
PRISM Licensed to Michigan State Police, DNA Unit GenotypeiQ 2.1
Ill‘IIIIIIIII'TI'IIII'TTIT‘IIIIIIIIYY'ITIIIII‘II'IIIIYVIIIIIII’IIIIII'III'I

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420

 

ll'lll'llDIIII'IOIIIII'III'Ill'lID'lll'lll'llI'tlllIIO'IIIIIIOIIII'OIIIO
80 100 120 140 150 180 200 220 240 260 280 300 320 340 360 380 400 420

A9—N—15(3UL) 4 Blue N-15(3UL)
600

00
200

 

A9—N-15(3UL) 4 Green N-15(3UL)
00
200

     

A9-N-15(3UL) 4 Yoiiow N-15(3UL)
200

100

 

A9—N-15(3UL) 4 Red . N«15(3UL)

111 111 111L133

 

 

153

 

ABIA Thesis data - concentrated amp. product
PRISM Licensed to Michigan State Police. DNA Unit Genotyparo 2.1
TYIYTI’TIII[YYfIlI—‘FFITIIttfifrIifiIiiilfttI'IITTTVVI'IIY'FII

80 100 120 140 160 180 200 220 240 260 280 300 320 340' 360

 

l'o ll. Iii 'III 'II III ii'i II' III 'lIl Iii O'Il I'I il| llll iii '0! l'l ii
80 100 120 140 160 180 200 220 240 260 280 300 320 340 360
C1 ~X-5(3UL) 15 BIUO X-5(3UL)

1500
1000
500

 

 

 

 

 

C1-X-5(3UL) 15 Green X-5(3UL)

-2000
~1500
-1 000

I l -500

 

 

 

 

cr-x-slaul.) 15 Yellow x-slam.)
been

r800

r400

7200
9.

 

 

     

C1 ~X-5(3UL) 1 5 Rod X-5(3UL)

11 - 1 111SEE

 

 

154

 

AB“; Thesis data - concentrated amp. product

PR1SM Licensed to Michigan State Police, DNA Unit Genotyperw 2.1
IIITr'rrIIIITTIIIIrT—rTII—rrTIIIIIIIIIITIIIIII'll'lll'l'jI'IrI

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

 

'III...IIIIO'Ili'IIIlIII'|IO|III!I."IIIIIIC'III"IIIIIO|OII
80 100 120 140 150 180 200 220 240 260 280 300 320 340 360
C5-X-15(3UL) 17 Blue X-15(3UL)

 

 

 

 

 

2000
1500
1000
l 500
I I - J A ,
m 1 6
2127' 1255]
13 1a
21 5 3
cs-x-15(3UL) 17 Green x-15(3UL)
2000

1500
. 1000
l ‘ .1 500
ii! IEJF IE! 1!!
1595: 1953 I!!! 704
ll!!!

 

 

 

 

 

   

v
EEEII
cs-x-15(3UL) 17 Yellow Xo15(3UL)
900
500
300
“.1
IE
(I!)
cs-x-15(3UL) 17 Red x-15(3UL)
800

 

 

00
00
200

 

 

 

 

155

 

ABIA_ thesis data - concentrated amp. product _
Licensed to Michigan State Police. DNA Unit Genotypero 2.1

T—rIIIYfiIT‘II'IIIIIIIIIIIrIrTITW’TrrTI'IT

200 220 240 260 280 300 320 340 360

 

TTTYT'IIIITIIIT

100 120 140 160 100

 

 

   

 

 

 

 

 

 

 

100 120 140 160 180 200 220 240 260 280 300 320 340 360
E5-W-15(3UL) 30 Blue W-15(3UL)
3000
2000
1000
3209 2201
18 23
1232
E5-W-15(3UL) 30 Green W-15(3UL)
2000
n 1000
[15
1209|
766
E5—W-15(3UL) 30 Yeflow W~15(3UL)
1500
1000

‘ j 600

11]
1322

 

ES-W-15(aUL) 30 Red w-15(3UL)

600
00
200

 

 

 

156

 

ABIA Thesis data - concentrated amp. product
Licensed to Michigan State Police, DNA Unit Genotypero 2.1
IIII'IIIjiffII’IIlrf"II—T'IIITWTIj—I‘IIIIjTrTWTIIrT'ITIIYTI'III

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

ll.ll'lll'lllllll'Illllll'lil'lll'Illllllllll'ltllIUIIIII'III
80 100 120 140 160 180 200 220 240 250 280 300 320 340 360
A5-AA-5(3UL) 4 Blue M-5(3UL)

 

 

 

 

 

 

 

   

 

 

 

 

600
600
400
200
A5.M»5(3UL) 4 Green AA-slauL)
400
L400
L ~200
RE]
30
A5oAA~5(3UL) 4 Yellow AA-5(3UL)
00
. 00
L 200
[I] :3 IE
EB BEE mm
111! IE
[IE
A5-AA-5(3UL) 4 Red AA-5(3UL)

 

11111 111

 

 

 

 

157

AB'A Thesis data - concentrated amp. product
PRISM Licensed to Michigan State Police, DNA Unit Genotyper® 2.1

TTIIIfi'I‘I—rTIII1YI—TIIII—[TTI]II‘V'ITTIYITlfiTrIIIIIIYrIII—rfT
100 120 140 160 180 200 220 240 260 280 300 320 340 380

 

1 III III III III III ill '1! 1'. II. III Ill 1'. III iii '1! ll. ll. lit III
100 120 140 160 180 200 220 240 260 280 300 320 340 360

E1oM-30(3UL) 28 Blue AA-30(3UL)

2000
1 500
1 000
500

 

 

 

1500
1000
500

E1-AA-30(3UL)

 

 

     

E1~AAo30(3UL) 28 Yellow AA-30(3UL)

1000
500
l l- 11 _ ,

E1-AA-30(3UL) 28 Rod AA~30(3UL)

 

 

600
00
200

 

 

 

 

158

AB'A Thesis data-concentrated amp. product
PRISM

Licensed to Michigan State Police, DNA Unit Genotypem 2.1
TlrIlI'rTIll ll—rWIIIIII[lrf’UYTlIII'rfT—IIIIITII‘IIIIIIIIT‘I

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

IIIIlllllil‘lllilll'lll‘lllllII'IDIIIII'IIIIIII'III'IOI'II.
100 120 140 160 180 200 220 240 260 280 300 320 340 360
F2-FF-30(3UL) 34 Blue FF-30(3UL)

3000
2000
. l 1000
1 _ e - ---A _
IE IE I18
m 3176

F2-FF-30l3UL) 34 Green FF-30(3UL)

 

 

r4000
-3000

LA! 92000
A] 71°°°
x 27|
4261‘ - 1779'
Y] 1 28
3597 1566

F2-FF-30(3UL) 34 Yellow FF-30(3UL)

l l J @233
%300

 

 

 

 

 

 

 

 

 

 

 

 

 

Ilfl iii 11
1087 1049'
m 12
700

 

F2-FF-30(3UL) emed FF-30(3UL)

E600
-400
~200
F

 

 

 

 

15S)

AB'A Thesis data - concentrated amp. product
PRISM

Licensed to Michigan State Police, DNA Unit Genotypenv 2.1
IvIrTIITrerI—FTTI'ITIII'TTIIIIf‘rrTI‘TTTiYTIIT—rlrleITTTIIVIVI

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

OIIIIII'IIIIIII'III'III'IIIIIII'IIIIIIO'IIIIIIIIIII'III'II

100 120 140 160 180 200 220 240 260 280 300 320 340 360
CQ-J-5(3UL) 19 Blue J-5(3UL)

 

 

   

 

   

 

000
600
400
200
m a!
719

Gee-slam) 19 Green J-5(3UL)
1500
1000
500

{394]
m
339

ce-aoslauu 19 Yellow J.5(3UL)
400
200

m
519 IE 551
E IE
EH

CNSGSUL) 19 Red J-S(3UL)

000

00
400
200

 

 

 

160

 

ABIA_ Thesis data- concentrated amp. product .
Licensed to Michigan State Pollce, DNA Unit Genotypero 2.1
1V,IIV'l"I‘ITTTTYYrVTTIVTTI'YUI'VTI'IIIIfiIjrrrI"IIYII[III‘IIIIYI

80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

so 100 120 140 160 130 200 220 240 260 230 300 320 340 360 380
82-M-5(3UL) 6 Blue M-5(3UL)

 

 

 

 

 

32146901.) 5 Green M-5(3UL)
00
300
200
100
E‘ m in E]?! [E
HE m [E BE
EB IE
[it
emu-slam.) 3 Yellow M-5(3UL)
200
150
100
so
82-M-5(3UL) a Red M-5(3UL)
000
00
200

 

 

161

 

thesis data - concentrated amp. product

ABIA
PRISM Licensed to Michigan State Police. DNA Unit

Genotypenb 2.1

 

lTjIr‘iI‘ijTIIT'IIIIIIIITII'IIIl‘TrlII'IIIIIYU

 

 

 

IIIIIIVIIr

 

 

 

 

 

 

100 120 140 160 180 200 220 240 260 200 300 320 340 360 380
IIIOI|III'IIII IIIIIII'II'I1|..1II'UIIIIIIIIII'II I'III'IIII
100 120 140 160 100 200 220 240 260 280 300 320 340 360 380
BS-O—S(3UL) a Blue O-SlSUL)
400
300
200
100
.AA A !— ‘L.
Be-o-slauL)
400
200
a I
Bs-o-slaul.) a Yellow O~5(3UL)
200
150
100
50
BS—O-5(3UL) a Red 05901;
600
400
200

 

 

162

 

AB'A Thesis data - concentrated amp. product
Licensed to Michigan State Police. DNA Unit Genotypero 2.1
IIIIIfirIIYTTrIII'TTI’I‘I’IrI—I—II]III'IITIITrlI—VIrfiTIIIrI

O 100 120 140 160 180 200 220 240 260 200 300 320 340 360

 

1|II'III'IIIIIIIlIIIIIII'Ill'lll'llI'llllIllllII'IIlIIII'

O 100 120 140 160 180 200 220 240 260 280 300 320 340 360
E9-U-15(3UL) 13 Blue U-15(3UL)

 

 

 

 

 

 

 

 

 

 

 

600
400
200
‘d '_A_ __ ‘ . LL- 3W _ ‘ ‘ A- L-.- “A . '
mil-I leg - a: m
739 E] gm [E HE a3
[Hi IE m m
IE 17 m
167 288 an
444 m
EO-U-15(3UL) 13 Green U-15(3UL)
1500
1000
500
14 m E
230
[E 247
m
E9-U-15(3UL) 13 Yellow U-15(3UL)
300
200
100

 

 

 

Es—u-lslauL) 13 Flea U-15(3Ul.)

#—

 

 

 

163

ABIA Thesis data - concentrated amp. product

PRISM Licensed to Michigan State Police, DNA Unit Genotypr 2.1
GIVIWIIIII'ITT'IIYIIr‘I'ITIT‘ITII—I—IIIII‘IIIITIIII‘III'VIIYI’IIII‘

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

Illll'lllllll'III'III'IIIIIIllilI'lll'Ill‘tltlIIIIOIIIOII‘
100 120 1‘0 160 180 200 220 240 250 280 300 320 340 360 380
B 1 0°U-30(3U L) 1 1 Blue U‘30(3U L)

 

 

600
400
; 1 200
LA. ‘ ._ - A__ 4...“ u u.
m an
[$25 [D Em rm
IE 54 IE
rm EB ma
161
200 676

BtO-U-30(3UL) 11 Green U—30(3UL)

900
600
300

 

 

 

 

B10-U-30(3UL) 11 Yellow U-30(3UL)

400
300
200
100

   

B10-U~30(3UL) 11 Red . U-30(3UL)

600
400
200

 

 

 

 

 

164

AB'A. Thesis data - concentrated amp. product
PR‘SM Licensed to Michigan State Police. DNA Unit Genotypero 2.1
Y1‘rIITIII—III'II'Ir‘Ij—IUII'TTTI—TllllIIIIIjIIII'iTV—TVIfrIII'II

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

Illllllllllllll‘lllllll'llllll‘lltt'ttlllll'lll’Ill'llt'lI
100 120 140 150 180 200 220 240 260 280 300 320 340 360
D10-Q-15l3UL) 26 Blue Q15(3UL)

 

 

 

 

 

1500
1000
J 500
[E m an E141

1079 IE3]

at: - E

. EB

DtO-Q-tS(3UL) 20 Green o-lslaul.)

000
000

2000
1000

: [E
m

D10—Q-15(3UL) 26 Yellow 0-15(3UL)

 

 

1000
500

 

 

 

El
1389
13

1116

 

 

 

 

Dto-G-15(3UL) 20 Bee o-lsalUL)

00
400
200

 

 

 

165

 

ABIA Thesis data - concentrated amp. product
PRISM Licensed to Michigan State Police, DNA Unit GenotyperQ 2.1
fijw'IIIIII—IIITrTTI'I[III'IIIIIIIITI—TITIIIIIT—rI—I—TirtrrtIT'AYI'

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

I I I I I I I‘| I I I ' I l I ' I I l i I I I I I I I ' I I I I I I I I I I I I I I I I I I I l I I I I I I I l l I
100 120 140 150 130 200 220 240 250 230 300 320 340 350
Bees-slam) 23 Blue BB»5(3UL)

 

 

 

 

 

   

 

 

1 500
1000
500
J 4 .
IE IE E11
m
an
DS—BB-5(GUI..) 23 Green BB—5(3UL)
900
00
300
1 51
256
IE
IE
DS-BB-S(3UL) 23 Yellow BB-S(3UL)
600
00
200
m
an W IE
IE
33
DG-BBFSGSUL) 23 Red BB-S(3UL)
600
00
200

 

 

166

 

AB'A_ Thesis data — concentrated amp. product
PRISM

Licensed to Michigan State Police, DNA Unit Genotyper® 2.1
I'II‘III'IIII‘TIIIIIIY—IITITI'rIIIrI—rmrmIIIITIIIIl'T—T'Y—Ift
100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

III'IIIIIII'III'III'III'IIIIIIl'III'III'IIIIIII'III'III'II

100 120 140 160 180 200 220 240 260 280 300 320 340 360
D4-L-15 22 Blue L-15'

 

 

     

 

 

 

1500
1000
500
D4-L-t'5l 22 Green L-15
2000
1000
A A AA
29] IE
479 423
30 Ilfl
534 415
D4-L-15 22 Yellow L-15
400
200

 

 

ll_l 3 IE
Ella-178

 

D4~L~1S

900
600
300

 

 

 

1(57

 

APPENDIX F

Electropherograms generated from ladders
and quality control samples.

168

ABIA Thesis data
PRISM

Licensed to Michigan State Police. DNA Unit Genotypem 2.1
IIl[TII'TIIIT‘IIIFTrTIIIIr‘IYTTIT‘rfiI‘rIlIrIfIITWTIjTVIIFD

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

IIII‘IIIIIIIIIIIllll'lll'Ill'III'III'III'II||III'III'III'
0 100 120 140 150 180 200 220 240 260 280 300 320 340 360
A3310 BLANK 3 BIUB 310 BLANK

80
60

A3—310 BLANK 3 Green 310 BLANK

 

A3—310 BLANK 3 Yellow 310 BLANK

3O
20
10

A3—310 BLANK 3 Bed 310 BLANK

1000
500

 

 

 

 

 

169

PRISM Licensed to Michigan State Police. DNA Unit GenotyperQ 2.1
ITWTIIT—r—rIfiI—rrllIIIIrTIW—rIITII1IrIrrI—III—III‘III—TIrrTTITr'IrI—fYTI'II
IO 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400

 

IIIIIIII'IIIIIIIIIII‘III‘IIIlIII'III'III'III'IIIIIII'III'III'III'll

IO 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400
A3—310 BLANK 381m 310 BLANK

100
50

A3—310 BLANK 3 Green 310 BLANK
40
20
l

A3310 BLANK 3 Yellow 310 BLANK

A3-310 BLANK 3 Red 310 BLANK

1 1111 - 111 1‘3?"

01

 

 

 

170

 

ASIA Thesis data
PRISM

Licensed to Michigan State Police, DNA Unit Genotypero 2.1
IIITr—IYjI'IlIl—r'IIIrITTTrrI[IT—rrIIYIIIIIIrrIIjII

100 120 140 160 180 200 220 240 260 280 300 320 340

 

III

I I | I I I | I I I I I I I | I I I ' I I I | I I I I I I I ' I I I I l I I | I l I I I I I I I I I I
100 120 140 160 180 200 220 240 260 280 300 320 340
A3—310 BLANK 3 Blue 310 BLANK

15
10

AIS-310 BLANK 3 Green 310 BLANK

20

I 'l I - ' . . p I : l ‘.I
I." ' l . .l I ‘l I ". t I

A3—310 BLANK 3 Yellow 310 BLANK
20

10

A3—310 BLANK 3 Red 310 BLANK

900
600
300

 

 

 

 

171

ABIA Thesis data
PRISM

 

 

Licensed to Michigan State Police. DNA Unit Genotypero 2.1
IIIIITfirfiITIIrIfIIUTIlll’I'IIIlf—ITII I—FIYII‘TT‘YIIII'IVIITI
100 120 140 160 180 200 220 240 260 280 300 320 340 360
III'IIIlIII'III'III'III'III'III'III'III'III'IIIlIIIIIII'l
100 120 140 160 180 200 220 240 260 280 300 320 340 360
A3-310 BLANK SBIue 310 BLANK
30
i 20
a 10
i i 1 1 1 1
A3310 BLANK SGreen 310 BLANK
80
60
0
20
A3-310 BLANK a Yellow 310 BLANK
O
20
A3—310 BLANK 3 Red 310 BLANK
900
600
300

 

 

 

172

 

APR§IS%A Thesis data

Licensed to Michigan State Police, DNA Unit Genotyperfi 2.1
T‘I—IITTI’I'II] TTI’rITIITV I‘ITI I'lij—I—IT—erU‘I—I—I [ITI] UTIIIT—I—[rlj'I

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

llIIIII'III'III'III'III'III'IIIIIII'III'III'IIIIIIIIIII'III'I
100 120 I40 160 180 200 220 240 260 280 300 320 340 350 380
A3310 BLANK 381UB 310 BLANK

No Size Data

A3-31O BLANK 3 Green 310 BLANK

No Size Data

A3—310 BLANK 3 Yellow 310 BLANK
20

15
10

. ,‘ l ‘, ‘ a g I - g . , , ‘
i. -. .. 1.. ‘ “ i 1
s . - . . - i

A3—310 BLANK 3 Red 310 BLANK

1000
500

 

 

 

 

 

 

173

ASIA Thesls data
PRISM

Licensed to Michigan State Police, DNA Unit Genotypem 2.1
ti‘jrjr'II[TITIYT—rri’l'lritj'f'rrTTT—I'Y—TIIVIlrleIfTrIr

O 100 120 140 160 180 200 220 240 260 280 300 320 340

 

’III'IIIIIII'IIIIIII'III[Ill'IlI|llI|lIIllIIlIII|III|II
O 100 120 140 160 180 200 220 240 260 280 300 320 340
A3-310 BLANK 3 Blue 310 BLANK

20
10

A3—310 BLANK 3 Green 310 BLANK

 

fl‘I :- ‘ ' 1 III I | HI ~. '1" 5

A3-310 BLANK 3 Yellow 310 BLANK
1 5

i . 1 0

A3310 BLANK 3 Red 310 BLANK
600

1 IIILI+1-J1‘233

 

 

 

 

174

ASIA Thesis data
PRISM Licensed to Michigan State Police, DNA Unit GenotyperO 2.1
YIfTitrIrl’IIvr'rrt—tfiitttjIV—rrfittlVIVITUTIlti’ltifi'

100 120 140 160 180 200 220 240 260 280 300 320 340

 

I.III'IIIIIIIIIII'IIIIIII'III.IIIIIIIIIII|IIIIIII'
100 120 140 160 180 200 220 240 260 280 300 320 340
A3-310 BLANK (3) 3 Blue 310 BLANK (3)

20

. 10
I l I
' . u i_|

A3—310 BLANK (3) 3 Green 310 BLANK (3)

20
15
' ' . I I r , 10
M3 ' -" I ' ._ . ' K ' ‘ .' :tL'; '. _ .l 9' ~.‘ 5

g .. "r

A3-310 BLANK (a) 3 Yellow 310 BLANK (a)

A3—310 BLANK (3) 3 Red 310 BLANK (3)

1000
500

 

 

 

 

175

. Licensed to Lynne Helton, Michigan State Police Genotyper® 2_1

ABIA Thesis data~contamination check
PRISM

 

IIIfiITITIY1I|VITIIl’T'I’rITTTU'rTj‘I’TTTY—IIIIIll'rrTIlIIUIIII'II'II‘I'I'IV

I) 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400

III.IIIIIIIIIIIIIII'III'IIIIIII'III'III'III‘IIIlIII‘III'IIIIIIIIIII'II

D 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400
A7'AMP BLANK (C) 5 Blue AMP BLANK (C)

40
20

A7-AMP BLANK (C) S Green AMP BLANK (C)

No Size Data

A7-AMP BLANK (C) 5 Yellow AMP BLANK (C)

No Size Data

A7-AMP BLANK (C) 5 Red AMP BLANK (C)

M11111 333

 

 

 

 

176

ASIA Thesis data - Group 1
PRISM

Licensed to Michigan State Police, DNA Unit Genotypem 2.1

 

L'lII'III'III'III’IIIIIII'III'III|III'III'IIIIIIIIIII'III'I
100 120 140 160 180 200 220 240 260 280 300 320 340 360 380
A5-AMP BLANK 4 Blue AMP BLANK

30
20
10

I v
| i
I . I . |

A5-AMP BLANK 4 Green AMP BLANK
30

20

1 I , ,- , I 1' H} . .. .' I .Y I ‘ ‘lilj'

i I 'i i '

AS-AMP BLANK 4 Yellow AMP BLANK 1 5

10

AS-AMP BLANK 4 Red AMP BLANK

600
7 400
200

 

 

 

 

177

ABIA Thesis data - Group 2
PRISM

Licensed to Michigan State Police, DNA Unit Genotypem 2.1
Y‘ITTTWII1IITIII’TjFI—I'YI‘II'Irr'I'II'IIUI'TII‘rIT‘IjIIITIf

100 120 140 160 180 200 220 240 260 280 300 320 340

 

DIIIIIIIIII'III'IIIIlll’IlI'Ill'IlI'IlI'IIIIIII'IIIIIII
100 120 140 160 180 200 220 240 260 280 300 320 340
AS-AMP BLANK (2) 4 Blue AMP BLANK (2)

4O
20
l "I I ' l I

AS‘AMP BLANK (2) 4Green AMP BLANK (2)

20

. 15

. _ i , t 10

, '1 I I- ," ‘1‘ ._._i 'l I . .l' 'l

.7" ‘-. ‘ . ,5.“ ‘ ffl 1‘ ‘(r ‘ .l l'. ' .. "lll" 5
A5-AMP BLANK (2) 4 Yellow AMP BLANK (2)

15

10

. " '. l . I ‘ .
. ', I i l . I- , ,It | . ,' f , , .: ‘ I , ,: 5
. I ".‘I '. ‘ l . It ,- -; | l 1' [till ; ‘ ’

AS-AMP BLANK (2) 4 Red AMP BLANK (2)
800

600
400
200

 

 

 

 

 

 

178

éBISfi thesis data - Group 3

' Licensed to Michigan State Police. DNA Unit GenotyperQ 2,1
TlIT'IYUUIrf'IrrjIUII—ITB']'ffi'frI'Ii'Vr'lIrV‘UU'l

100 120 I40 160 180 200 220 240 260 280 300 320 340

 

u I I u I I o a I I I n . I I I I | 0 I I I I I I I I I I I I n l I l I I I I o I I o I I ' I i I I
100 120 140 150 180 200 220 240 260 280 300 320 340
AS-AMP. BLANK (3) 4 Blue AMP. BLANK (3)

4o
30‘
20
l t ’ i 10

' '.l , l . - -_ , . '

A5-AMP. BLANK (a) 4Green AMP. BLANK (3)

20

p 15

. I. p I; ‘ . ‘ . lo

.‘ “3| ' l l I :III ‘1." ' ' 1'

 

As-AMP. BLANK (a) 4Yellow AMP. BLANK (a)

. l I I ' II
.. . - l ' I' I "

A5-AMP. BLANK (3) 4 Bed AMP. BLANK (3)
800
00
400
200

 

 

 

 

179

AB'A Thesis data - Group 1 .
PR'SM Licensed to Michigan State Police, DNA Uni! Gonotyporo 2.1
II'7'!lililrrlllltf'lr—rllVIIIUUU‘IIIIIrTjUUrr‘TTIl’IUYIVVt'I

00 120 140 160 180 200 220 240 260 280 300 320 340 360 380

 

iIIIIIIIIII.IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
100 120 140 150 180 200 220 240 260 280 300 320 340 350 380
AWMKXIJOONNNJ. 58MB “KKXJCONTROL

A7—WOOD CONTROL 5 Gwen WOOD CDNTROL

.'| '._‘ i 1| . . 4‘ , I . | , .I -

200
150
100
50

800
800

00
200

 

 

180

 

 

 

A
1%:ng

Thesis date - Group 1
Licensed to Michigan State Police. DNA Unit

GenotyperO 2.1

 

I I

'U‘II‘l—T'UIV'YIYITI‘YTITWYIl'fI‘IfiIUijjjfrliiltlrj

 

 

 

 

 

 

 

 

 

 

 

 

 

 

100 120 140 160 180 200 220 240 260 280 300 320 340
IIIIIIIIIII'III'OOUII.I"IIIIUII'IIII'IIlIlI'I'I'IU'I
100 120 140 160 180 200 220 240 260 280 300 320 340
E1-MB-ILC 273MB MB-ILC
1000
I l 500
Ill
1464 an
“E
EIII
Ei-MB-iLC 27 Gwen MB—lLC
1500
1000
500
E! [[3 I15 HE} ([1
112! ifiiiiifli 1333 [2:1
EI-MB-ILC 27 Yellow MBv-ILC
' 400
300
200
100
Ei-MB-iLC 278M MB'ILC
800
00
on
200

 

 

181

ABIA
PRISM

Thesis data - contamination study

Licensed to Michigan State Police, DNA Unit

Genotypem 2.1

 

l’ I’ I f'ffil I If I Y I Yrj’j’fi Y "rj rT—l' l' l' T I r I I

rfijfi I T—rrl I I I I

 

 

 

 

 

 

 

too 120 140 160 180 200 220 240 260 280 300
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
too 120 140 160 180 200 220 240 260 280 300
012—M8 (C) 26 Blue M8 (C)
400
I I M 200
IE 33
237 257
IE E
813 232
th-MB(C) 266mn MB (C)
600
00
200
IE1 IE1 in HE IE ID
734 292 a 240 um
Die-Ma (C) 26 Yellow MB (C)
200
150
100
50
Ill IE 11
204| [m 129]
12 m 117
me we
D12-M8 (C) 26 Red M8 (C)
900
600
300

 

 

 

 

182

 

ABIA Thesis data - contamination check
PRISM

 

 

 

          

 

 

 

 

Licensed to Michigan State Police, DNA Unit Genotypero 2,1
Il‘l’firlTT—‘IljlrlI[jIfirili[UTTITIW1TVI‘Till1U—I
100 120 140 160 180 200 220 240 260 280 300
IIIIIIIIIIIIIIIIIIIIIIIIIIIII'IIIIIIIIIIIIIII
100 120 140 160 180 200 220 240 260 280 300
E3-+CONTROL (C) 28 Blue +OONTROL (C)
1000
A I 500
32
E3»+CONTROL (C) 28 GNOfl +OONTROL (C)
1500
1000
500
E [E E IE [E
m 1407 EE (ES
E3+OON1ROL (C) 28 Yellow +CONTFiOL (C)
@900
9600
I I 9300
m m ng
m 749
II]
E
Es-KJONTROL (C) 28 Red +OONTHOL (C)
. Leon
54500
{-400
7200

 

 

 

 

 

183

 

ABIA_ Thesis data - Group 1
PRISM Licensed to Michigan State Police. DNA Unit Genotypem 2.1
IFIrrtt'l’lTTIlrrr‘IVIIIIIYTTIIITrTIIrY'fiTfirVIIITT‘IT'II’I’IIII’f'III

0 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIiIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
50 30 100 120 140 150 180 200 220 240 260 280 300 320 340 360
E3-+ CONTROL 28 Blue +OONTROL

 

 

 

 

 

 

 

 

 

—1ooo
—soo
E3—+ CONTROL 28 Green + CONTROL
2000
1500
1000
AI I47 500
E3 IE3 Ell (E! II]
1 5 5 6 EH 7 3 3
53+ CONTROL 28 Yeliow + CONTROL
1000
500
II] III II;
flfiii 1014 E!!!
H“
III]

E3-+ CONTROL 28 Red + CONTROL

800
00
400
200
#4 A _

 

 

 

 

184

 

QBIA Thesis data - Group 2

 

Licensed to Michigan State Police, DNA Unit Genotypens 2.1
I I I r‘T I I r r't t t T r I I I 1 l’ I I r‘rfi r I I I rtfi I I I I I r I I r I I 1 U
100 120 140 160 180 200 220 240 260 280 300
I I I I I I I I I I I l I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
100 120 I40 160 180 200 220 240 260 280 300

E5-+CONTROL (2) 29 Blue «CONTROL (2)

 

 

 

E5-+CONTROL (2) 29 Green +CONTROL (2)

 

 

 

 

 

1000
see
IKE mm [EB EH3
£5-+CONTROL (2) 29 Yellow +CONTROL (2)
L800
9400
M P200
P
III II] [£1
2:! 507 £12!
[I]

ES-+OONTROL (2) 29 Fled +CONTROL (2)

' I900
i800
r300

 

 

 

 

185

 

ABIA Thesis data - gmup 3
IDFEESIVT

 

 

Licensed to Michigan State Police. DNA Unit Genotypero 2.1
T “F I’ l' I I T I i I T T r r r I I'— l I I I I I I ‘ ‘I’ Y 1"1' 1 I I I I Y I l I l’ T I I I I
100 120 140 160 180 200 220 240 260 280 300
I ' I I I l I I I I I I I I I I I I I I I ' I I I ' I I I | I I I | I I I | I I I I I I I
100 120 140 160 180 200 220 240 260 280 300
E1-+CONTHOL (3) 27 Blue +CONTROL (3)
2000
‘AJ[ 1000
'
E!!!

   

E1-+CONTROL(3) 27 Green +CONTROL'(3)

4000

fl 2000
E3 ‘13 El] II]
EEEEI [£521 lain! 155‘ fliflfi

E1o+OONTROL (a) 27 YoIIow +CONTROL (3)

 

 

 

 

2000
h I! 1000
III III I]!
fiflifll 2371 {155
III
EEII

E1—+CONTROL (3) 27 Red «tmNTROL (3)

900
600
300

 

 

 

1863

 

Thesis data
Licensed to Michigan State Police. DNA Unit

ABIA
PRISM

100 120 140 160 180 200

220 240 260 280 300 320 340

GenotyperO 2.1

360

100 120 140 160 180 200
A1-LADDEH-3 15 8103 LADDER

220 240 260 280 300 320 340

1 21 24 2 3
21 33 2 2 2
1 2 2 2
391 33 19 25
29 21
27 41
23 2
23

AIcLADDER-a 15 Green LADDER

24. 2 31 3
462 28. 3 3
2 77 32. 3 1 51
73 2 40 47 54 301351 2
2 29. 3 34 10. 1 2 34 21
53 3 33. 35 33 24 36
2 30. 61 11 14. 19 29
678 40 38 46 19
27. 31. 46 73 31
76 353 35. 1
107 501 27
831 62
46 32
42 34.
46
67

187

360

 

ABIA Thesis data
PRISM

Licensed to Michigan Siale Police, DNA Unit

 

 

A1-LADDER-3 9 Yellow LADDER

300
200
100

 

 

A1-LADDER-3 9 Red lADDER

Lilli 11-11.33?

 

 

188

ABIA. Thesis data
PRISM

Licensed to Michigan State Police. DNA Unit Genotypem 2.1

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380

IIII‘III|III|III'III|IIIIIII'IIIIIIIIIII‘III'III’III‘III'
100 120 140 180 180 200 220 240 260 280 300 320 340 350 380
A1-LADDER‘3 9 8100 LADDER

A1-LADDEH-3

 

189

ABI;- Thesis data
PRISM

Licensed to Michigan State Police. DNA Unit GonotyporO 2.1

A1-LADDER-3 15 Yolow LADDER
300
200
100

 

 

 

 

 

 

A1-LADDER-3 15 Rod LADDER

LDUL A i- i- i 333

 

 

 

 

190

 

QEISIAM' Thesis date

Licensed to Michigan State Police, DNA Unit Genotypeto 2.1

100 120 140 160 180 200 220 240 260 260 300 320 340 360 380

100 120 140 160 180 200 220 240 260 260 300 '320 340 360 380
Ai-LADDER-a 10 Blue LADDER

At -LADDER-3 10 Green LADDER

24. 2 3

454 2 31
2 768 38
73 81 3
2 45 6

3 3
3 3
3
11
O 35

54 3 32

2 45

65 391

27. 34
6 31.

62
7
45
47

104 49
28. 33.
29. 34

42

30.
65
32.

191

 

ABIA Thesis date
PRISM

Licensed to Michigan State Police, DNA Unit Genotypero 2.1

 

A1-LADDER-3 10 Yellow LADDER

 

 

 

 

300
200
100
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341 11 14 312 12 15 253 9 12 1s
a 317 258 9 221 7 1 13
374 ‘27s 16 254 13 214 166 194
332, 13 10 217 233 191
10 31s 21s 1239 243 214
333 231 277 227 211

 

 

fifiifl 232

A1-LADDERo3 10 Rod LADDER

111 1 - 1- 1L1???

 

 

 

1192

$394” Thesis date

Licensed to Michigan State Police. DNA Unit Genotypem 2.1
‘I’II'IIIIIIrIIIYIIII'I IIIIVTrIIIT‘TW—U[I—YVTIIF—rrIUTUV—I—IlVIII

100 120 140 160 180 200 220 240 260 280 300 320 340 360

 

III‘III'III'IIIIIII'III.III'III.III|III|IIIIIII'IIIIIII‘I
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mucosa 1 Green LADDER
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9300
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193

 

ABIA Thesis data
PRiSM

Licensed to Michigan State Police, DNA Unit Genotypem 2.1
A1-LADDEFi-4 29 Yeiiow LADDER

A1-LADDER-4 29 Red LADDER

 

194

 

ABIA_ Thesis data
PRISM Licensed to Michigan State Police. DNA Unit Genotypene 2.1
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2 301 21
28 4 241
23 24
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A! -LADDER4 29 Green LADDER

 

195

 

 

 

 

ABIA Thesis data
PRISM

. Licensed to Lynne Helton, Michigan State Police

 

 

 

 

 

 

 

 

At—LADDER tYeitow LADDER
300
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400
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196

ABIA Thesis data
' PRISM

Licensed to Lynne Hench, Michigan State Poiice Genotypeio 2.1

100 120 140 160 180 200 220 240 260 280 300 320 340 360

illlutlItl'nncltnillnI'ItojfluutuclionoluII|I0I|0|IIII||u
100 '20 ‘40 160 150 200 220' 240 260 280 300 320 340 360
A1-LADDER 1 Blu. LADDER

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88
50 63
44 34
481 401
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197

 

ABIA Thesis data

 

 

 

 

 

 

 

 

 

 

 

 

PRISM' Licensed to Michigan State Police. DNA Unit
Ai-lADDER 1Yeiiow LADDER
. 300
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198

ABIA. Thesis data
PRISM

Licensed to Michigan Stats Police. DNA Unit Genotypow 2.1
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0 100 120 140 160 180 200 220 240 260 280 300 320 340
AI-LADDER 1 Blue LADDER

800
600
400
200

 

 

A1-LADDER IGmn LADDER

 

21.115223”
51.125523.

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199

ABI; Thesis data
PRISM

Licensed to Michigan State Police. DNA Unit GenotyperO 2.1

At-LADDER 1 Yellow LADDER

A1 MDDER 1 Red LADDER

 

200