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VNTR Human DNA Using PCR
VNTR Human DNA Using PCR
VNTR Human DNA Using PCR
Using PCR
CHE 447-01
By Grant Akalonu
Grant Akalonu VNTR Human DNA Typing Using PCR
Objective/Introduction
evidence from a crime scene to suspects and victims, in a qualitative manner. DNA fingerprints
for specific individuals are determined by analyzing regions of chromosomes that vary greatly
among individuals These varying regions in chromosomal sequences are known as DNA
polymorphs. Analysis of polymorphic regions of DNA are also used to determine family
DNA fingerprinting first requires collection of hair, or bodily fluids from a crime scene
or victim. The sample is then stained and treated with a detergent to rupture the cell membranes
to acquire the DNA. The polymerase chain reaction (PCR) is utilized to amplify and examine
DNA regions of high polymorphic character. Polymorphic DNA regions fall into two categories,
VNTR (Variable Number of Tandem Repeats), and STR (Short Tandem Repeats). In this
experiment, the VNTR of interest is D1S80, which exists on chromosome 1 and has a 16
nucleotide sequence that is repeated anywhere between 16 and 40 times. After the DNA was
amplified at the D1S80 locus by PCR, it was visualized on agarose gel for analysis. The number
of bands on the the agarose gel indicates whether the student is heterozygous (two bands) or
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Grant Akalonu VNTR Human DNA Typing Using PCR
First, students rinsed their mouths vigorously for 60 seconds with 10 mL of saline water.
Then, the rinsed saline solution was swirled in a cup to suspend the cells and transferred to a a
centrifuge tube. The solution was then centrifuged at 800 rpm for two minutes. This was
repeated a number of times to create a cell pellet large enough for the next part of the procedure.
Then, the supernatant liquid above the pellet was removed and 140 L of lyse buffer was added
to the pellet. The mixture was then vortexed and incubated in a water bath at 55C for 15
minutes. The mixture was vortexed again and then incubated at 99C for 15 minutes. Next the
solution was centrifuged again for 2 minutes at 6000 rpm. 80 L of the supernatant was collected
and store on ice. Then, 20 L of primer, 5 L of the extracted DNA and a PCR Edvobead was
added to a 0.2 mL PCR tube. A similar PCR tube was prepared for the control DNA. and
another students DNA. This solution was mixed to ensure the dissolution of the bead, and then
quick centrifuged. The DNA samples were then amplified using the PCR, and its settings are
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Grant Akalonu VNTR Human DNA Typing Using PCR
Denaturation 94 30
Annealing 65 30
Extension 72 30
After the PCR is complete, 5L of 10x gel loading solution was added to the sample. The
gel was prepared by another group of students at the beginning of the lab period. 50x TAE
electrophoresis buffer was diluted to 1x, and 1.5% agarose gel was prepared by mixing 0.38g of
loaded into the gel after it solidified and ran at 140V for 45 minutes. The gel was then stained
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Grant Akalonu VNTR Human DNA Typing Using PCR
Results
2 (Control) 200
The human fingerprinting method described in this experiment was expected to show
homozygous to the DS180 genotype. However, a malfunction with the PCR machine did not
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Grant Akalonu VNTR Human DNA Typing Using PCR
allow for the visualization of different bands among students, as all of the gels in the class had
the same bands for every student, representing 200 base pairs.
1) There are two bands on the agarose gel from my DNA sample, which indicate that I
am heterozygous for the D1S80 genotype. It appears that George is also heterozygous
for the same genotype, as indicated by his bands at 200 and 600 base pairs. This
similarity in our results could possibly indicate a common ancestry between us, or
individuals. The main characteristic of these chromosomal regions that vary among
individuals is their length. The amount of base pairs that make up polymorphic DNA
polymorphic regions of DNA can be used to match samples of DNA from various
sources to an individual.
3) CODIS is the acronym for the Combined DNA Index System. It is a computer
system containing DNA fingerprints from crime scenes. It enables federal, state, and
local forensic laboratories to exchange and compare DNA profiles electronically,
thereby linking serial violent crimes to each other and to known offenders.
4) An short tandem repeat (STR) is a DNA sequence that varies from person to person
and is 2-4 base pairs in length. A variable number tandem repeat (VNTR) is a DNA
sequence varying from person to person but ranges from 15-70 base pairs in length.
Short tandem repeats are preferred in law enforcement, because they require a smaller
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Grant Akalonu VNTR Human DNA Typing Using PCR
amount of template DNA. This allows law enforcement to obtain viable experimental