Forensic DNA
Analysis
Mr. Kurunathan
Forensic Sciences
FAS
DNA Is In All of Your Cells
Body Fluids and Tissues
Blood
Semen
Saliva
Perspiration
Tissue
Bones and Teeth
Hair (if there is tissue on the root)
Urine, Feces, Vomit
Where DNA Evidence is Found
What is DNA?
DNA is the chemical substance which makes up our
chromosomes and controls all inheritable traits (eye,
hair and skin color)
DNA is different for every individual except identical
twins
DNA is found in all cells with a nucleus (white blood
cells, soft tissue cells, bone cells, hair root cells and
spermatozoa)
Half of an individual’s DNA/chromosomes come from
the father & the other half from the mother.
DNA Review:
DNA is a double-stranded molecule.
The DNA strands are made of four different building blocks.
An individual’s DNA remains the same throughout life.
In specific regions on a DNA strand each person has a unique
sequence of DNA or genetic code.
Brief History of DNA - (1985)
Multilocus
RFLP
Detects VNTRs:
Variable Number of
Tandem Repeats
Pure sample are
required
Brief History of DNA (Late 80s, Early 90s)
Single locus RFLP-
small
amount/degraded
sample
D2S44 probe
Lanes 6 and 10 match
Lanes 8 and 11 match
Restriction Fragment Length Polymorphism
Because of a change in DNA, restriction enzyme site is
created or lost
Measurements taken of fragments that vary in length
across people (length polymorphism) because they contain
VNTRs
can produce extremely low random match probabilities
Brief History of DNA (Early 90s)
PCR Strips (DQ alpha)
6 Alleles:
1.1, 1.2, 1.3
2, 3 or 4
A person can
have one or
two of these
numbers.
Two main types (90s - Present):
Variable Number Tandem Repeats (VNTR), Short Tandem Repeats (STRs)
Individual identification possible
Samples: Blood stains, semen
Mitochondrial DNA
Used in cases of severely degraded DNA
Individual identification not possible
Samples: Bones, hair shafts
Repeated DNA Sequences
Variable Number Tandem Repeats
(VNTRs) with a core sequence of 20-29
bps. -RFLP
Short tandem repeats (STRs) with a core
sequence of 2-5 bps. –PCR
Across the population, different
individuals have different numbers of
these repeats.
Short Tandem Repeats (STRs)
Currently the most used of all forensic markers
Individual identification possible
People differ in length at these loci
Each person has some STRs that were inherited from mother
and some from father
No person has STRs that are identical to those of either parent
Are located in the nuclear DNA (chromosomes)
Short Tandem Repeats (STRs)
Person 1 ..GCCAGCTAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5 6
Person 2 ..GCCAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5
Person 3 ..GCCAGCTAGCTAGCTAGCTAGCTAGCTAGCTT..
1 2 3 4 5 6 7
Short Tandem Repeats (STRs)
Locus :
Refers to the location on the chromosome.
Allele:
Refers to the type of DNA.
For STRs,
the allele will be the number of repeats.
Example:
Locus: D5S818
Alleles: 7,9
Paternal chromosome 5
CCAGATAGATAGATAGATAGATAGATAGATCC
Maternal chromosome 5
CCAGATAGATAGATAGATAGATAGATAGATAGATAGATCC
13 loci used in CODIS
STR Marker Chromosome Repeat Sequence Repeat units Other Alleles
TP OX 2 AATG 6 - 14
CS F1P O 5 AGAT 6 - 15 10.3
D5S 818 5 AGAT 7 -15
D7S 820 7 GATA 6 - 14
D8S 1179 8 TATC 8 - 19
D13S 317 13 TATC 7 - 15
D16S 539 16 GATA 5, 8 - 15
D3S 1358 3 TCTA* 9, 11- 20 15.2, 16.2
FGA 4 CTTT* 15 - 30 16.2 -30.2
22.3, 34.2, 46.2
TH01 11 AATG* 3, 5 - 12 8.3, 9.3, 10.3, 13.3
VWA 12 TCTA* 11 - 22 15.2
D18S 51 18 AGAA* 8 - 27 13.2, 14.2, 15.2
17.2, 19.2
D21S 11 21 TCTA* 24 - 38 24.2 - 35.2
Basic Steps in Analysis
Extraction:
Separates DNA from sample
Amplification or PCR:
Amplifies small portions of DNA (STR regions)
Separation:
Separates amplified fragments according to size.
FMBIO
Separates and Measures Amplified DNA
Color image of gel
Gel Electrophoresis
Black and white
image of STR gel.
Samples will have
one or two bands at
each loci.
ABI 310 Genetic Analyzer
Separates and Measures Amplified DNA
Sample will have one or two peaks at each loci.
Compare to a ladder that has all peaks at
each loci.
DNA Profiles are compared
TPOX CSF1PO D5S818 D8S1179
Blood stain 7,9 10,13 7,15 8,8
Suspect 1 8,9 10,10 9,10 11,12
Suspect 2 10,11 9,13 8,14 9,12
Suspect 3 7,9 10,13 7,15 8,8
DNA Profiles are compared
D8S1179 TPOX CSF1PO D5S818
Blood stain 7,9 10,13 7,15 8,8
Suspect 1 8,9 10,10 9,10 11,12
Suspect 2 10,11 9,13 8,14 9,12
Suspect 3 7,9 10,13 7,15 8,8
Mitochondrial DNA
Double Helix Chromosomes Ring of DNA
YES NO YES
The mitochondria has chromosomes that were only inherited
from the mother (MtDNA)
Mitochondrial DNA is only
16,569 letters long
[compared to 3 billion in
nuclear DNA]
There is a 900 base pair
region with a 1.7%
difference [D loop]
Nuclear DNA vs. Mitochondrial DNA
Double Helix Double Helix
46 Chromosomes One Ring
Multiple copies in each
One copy per cell mitochondria
Multiple mitochondria in
each cell
Nuclear DNA: Length is measured
mtDNA: Sequence is examined
MtDNA used for old or degraded samples
Basic Steps in Analysis:
Extraction:
Separates DNA from sample
Amplification or PCR:
Amplifies small portions of DNA
Sequencing:
Sequence of letters for amplified fragments
DNA Sequences are compared
Hair Sample AGCTAGATCGTTATTCCGAG
Victim AGCTAGATCGTTATTCCGAG
Hair may have come from the victim.
DNA Sequences are compared
Hair Sample AGCTAGATTGTTATTCCGAG
Victim AGCTAGATCGTTATTCCGAG
Hair did not come from the victim.
DNA Sequences are compared
Cigarette AGCTAGATTGTTATTCCGAG
Suspect #1 AGCTAGATCGTTATTCCGAG
Suspect #2 AGCTAGATTGTTATTCCGAG
Suspect #3 AGCTTGATTGTTATTCCGAG
Suspect #4 AGCTAGATTGTTATTCCGAG
Cigarette could be from Suspect #2, Suspect #4 or other
person with the same sequence.
Y-STR
Y- STRs are STR found on the male-specific Y Chromosome.
The Y-STRs are polymorphic among unrelated males and are
inherited through the paternal line with little change through
generations.
