Nicholas Henlon

December 15, 2014

Period 4A
Forensic Science
DNA profiling is a method used to identify a person by their DNA. This method does not analyze the
complete genome, rather it looks for repeats in the DNA sequence called tandem repeats. This method
can be used in identification, paternity tests, and DNA comparison. This method requires a sizeable
amount of DNA which can be acquired through PCR.
One method of DNA profiling is gel electrophoresis. Gel electrophoresis involves the use of a porous gel,
usually agarose, to help separate DNA based on its length. The first step of this process is to acquire DNA
and splice the DNA through the use of a restrictive enzyme. This splices the DNA by identifying a specific
sequence of DNA, usually a palindrome, and cuts it at that site. The agarose gel is made from combining
agarose, water, and buffer. This solution is then heated until it is thoroughly dissolved and is cooled. The
agarose gel is then put into a gel tray and cools until it is solid. While it is cooling, a well comb to form
wells for the DNA. The gel is then placed into the electrophoresis box and buffer solution is added to the
box until it is completely level. The DNA sample is then added to the wells. An electrical current is then
added to the electrophoresis box and the DNA moves toward the positive end, the cathode, because
DNA has a negative charge. After the electrophoresis is complete, ethidium bromide is added to the gel
and the fragments are viewed under a UV light. The DNA fragments are assorted by their length. The
lengths are dependent on how the restriction enzyme cuts the DNA and the shortest fragments will
travel the farthest.
The method used during class was different from what happens in the lab. One difference is that real
DNA was not used; instead, dye was used because it allows for the bands to be visible. If dye was not
used, the fragments would not be visible. Another thing that was done differently was that the DNA was
added to the gel before adding the buffer. This allowed for an easier insertion of DNA. Another
difference is that ethidium bromide was not used since the DNA was already visible.
Some advantages of gel electrophoresis are that it is relatively inexpensive and is a quick test. It can also
be used for quick comparison among two samples of DNA based on the bands in the samples. One
disadvantage is that it cannot show individual differences in DNA such as differences in DNA sequences.
Another disadvantage is that multiple tests will be needed to determine if an individual matches with a
DNA sample.
One example of new DNA technology is DNA microarray assays. This assay contains small amounts of a
large number of single-stranded DNA fragments which represent different genes. These DNA fragments
are placed in tightly spaced grids. This test is used to determine if hybridization occurred in the DNA of
the organism. This also tests for expression of various genes. This technology can be used to determine
if certain diseases such as cancer can be more likely to occur within an individual
This technology works by first isolating mRNA from a tissue sample. Reverse transcriptase is then used
to produce cDNA with the addition of fluorescently labeled nucleotides. The cDNA is then added to a
microarray which has single-stranded DNA. Excess cDNA is rinsed off and the microarrays that hybridize
show a florescent spot which varies based on the level of expression.