Elijah Meador 1

Gel Electrophoresis and Analysis of D1S80 Locus

Present in Student Population: 2PM lab section

Introduction
Tandem repeats are end to end duplications of a DNA sequence at a specific locus.
VNTRs or variable number of tandem repeats also called minisatellites are tandem repeats that
have a range from 7-100 base pairs that are commonly found in the euchromatin regions of
chromosomes. STRs or short random repeats also called microsatellites are arrays of repeats
shorter than 5 base pairs and are found in the euchromatin in plants, animals, and insect
chromosomes. The usefulness of VNTRs and STRs are that they are used in applications to
identify individuals because every individuals DNA fingerprint is different. (1)
PCR or polymerase chain reactions are used to amplify copies of a DNA fragment to
create many more copies of a particular DNA sequence. In the presence of a buffer, cofactors,
and dNTPs, a polymerase extends the primer sequences. This process amplifies the region of
interest and can be very useful in produce a larger sample of DNA from a small sample. (2)
DS180 contains a variable number of tandem repeats and is located on the chromosome.
There are several facts about the DS180 locus: More than 80% of all populations are
heterozygous, repeats are 16 nucleotides long, DS180 PCR product with zero repeat units is 142
BP, Every repeat will add 16 BP to the to the VNTR, PCR products range from 430 to 814 BP
long, 41 repeated units have been observed in the largest allele. (3)

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Materials and Methods

We used the thermocycler to house the PCR that copies the D1S80 locus from each
individual's DNA. Individual DNA isolation procedures were done to isolate cheek cells
suspended in saline solution by several microcenterfuge cycles and boiling the cell suspension
and chelex at 99C for 10 minutes and storing the isolated DNA on ice until the PCR reaction.
The PCR is used to amplify small amounts of DNA then the amplified DNA is delineated by
primers. There were two primers used to generate the PCR product that is much larger than the
repeat region. These two primers are D1S80 forward primer (5'GAAACTGGCCTCCAAACACTGCCCGCCG) and D1S80 reverse primer (5'GTCTTGTTGGAGATGCACGTGCCCCTTGC).
After calculating the specific volumes (in uL) for the components of the PCR reaction.
The components: 2x PCR mix (12.5 uL), 5uM primer 1 (1 uL), 5uM primer 2 (5uL), template
DNA (3uL), and water (7.5uL). A total volume of 25uL that enters into the PCR reaction cycle.
The PCR reactions are cycled in a 95C, four minute denaturalization step, 30 cycles of the
following; 95C 30 second denaturalization, 65C 30 second annealing, 72C two minute
elongation, and 35 cycles of 72C seven minute final elongation.
After undergoing PCR, electrophoresis samples were prepared by mixing the PCR
products with a loading dye. PstI electrophoresis marker was loaded into the first lane of each
gel. Samples were then loaded into the wells of the agarose (1.5%) gel. The electrophoresis
apparatus was then connected to the leads, and a voltage was applied. After undergoing
electrophoresis, the gels were visualized using UV light and analyzed

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Results
FIGURE 1

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Figure 1. Images of each gell were captured under UV light. All gels contained PstI DNA marker in the first lane,
and corresponding base pair values for each band are listed alongside the image for gel 1, other lanes contained
individual dna samples from the 2pm lab section.

FIGURE 2
Individual name

Sexton
Wingfield
Conway
Cook
Clark
Fisher-Hewett
Hinkle
Lee
Blevins
Koko
Marshall
Stowers
Strickland
Hunt
Pokuaa
George
Gyamfi
Wardell
Warren
Meador
Demery
Sentissi
Jones
Harris

Heterozygous or

DNA fragment/s

Number of

Homozygous

size/s (bp)

repeats in alleles

Heterozygous
Homozygous
Heterozygous
Homozygous
Heterozygous
Homozygous
Homozygous
Heterozygous
Heterozygous
Heterozygous
Homozygous
Homozygous
Heterozygous
Heterozygous
N/A
N/A
Heterozygous
Heterozygous
Heterozygous
Heterozygous
Heterozygous
Heterozygous
N/A
Heterozygous

659, 531
531
531, 435
659
659, 531
531
531
531, 435
531, 435
659, 531
531, 435
531
531
755, 531
No visible bands
No visible bands
531, 435
659, 435
531, 435
659, 435
531, 435
659, 531
No visible bands
659, 531

present
32, 24
24
24, 18
32
32, 24
24
24
24, 18
24, 18
32, 24
24, 18
24
24
38, 24
N/A
N/A
24, 18
32, 18
24, 18
32, 18
24, 18
32, 24
N/A
32, 24

Figure 2 data for D1S80 alleles present in the 2pm lab section. Figure shows homozygous or heterozygous presence
of the locus, visible band sizes, and number of repeats

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FIGURE 3
Allele (# of repeats)

Number of D1S80 allele in 2

% of D1S80 allele in 2 pm

18
24
32
38

pm lab section
9
18
8
1

lab section
25
50
22
3

Figure 3. Data for the D1S80 alleles shows allele frequencies in the 2PM lab section.

Discussion
VNTRs are useful for crime scene investigations and forensic science and the D1S80
locus in particular can be analyzed to determine someones DNA fingerprint. However, analyzing
this locus alone would not be enough information to prove a DNA fingerprint. Normally five to
six loci are amplified and analyzed to determine someones DNA fingerprint. One locus will not
typically provide enough variability to distinguish one individuals DNA from another. Only four
different alleles of the D1S80 locus seem to be present. The resulting data can be used to detect
the presence of certain alleles, such as eye color or hair color, but is not detailed enough for us to
determine an individuals DNA fingerprint based on these results. Much more data is needed in
order to correctly fingerprint an individual.

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References
(1)"Background." Department of Chemistry [FSU]. N.p., n.d. Web. 22 Feb. 2016.
(2) "Variable Number Tandem Repeat." Wikipedia, the Free Encyclopedia. Wikimedia
Foundation, Inc, n.d. Web. 22 Feb. 2016.
(3)"Variable Number Tandem Repeats." Department of Biology - College of Arts and Sciences.
N.p., n.d. Web. 22 Feb. 2016.