Anuththara Gunatilaka - Biology

VNTRs and DNA fingerprinting

A tandem repeat is a short sequence of DNA that is repeated at a specific
chromosomal locus. Tandem repeats are scattered throughout the human
genome. Some sequences are found at only one location, a single locus, in
the human genome. The number of repeated units vary between different
people for many tandem repeats. Such loci are termed VNTRs. One VNTR
in humans is a 17 bp sequence of DNA repeated between 70 and 450
times in the genome. The total number of base pairs at this locus could
vary from 1190 to 7650.
When profiles from a single VNTR locus from unrelated individuals are
compared, the profiles are usually different. However it is possible for two
individuals to have the same profile at one or two loci. But the chance of
more than one person having the same DNA profile at 4, 5, or 6 different
VNTR loci is very unlikely. When DNA profiles are used for forensic
purposes (e.g. DNA fingerprinting) 4-6 different VNTR loci are analysed.
((UOA), 2004)

Polymerase Chain Reaction

Sometimes called "molecular photocopying," a polymerase chain reaction
(PCR) is a fast and inexpensive technique used to copy small segments of
DNA. Because significant amounts of a sample of DNA are necessary for
molecular and genetic analyses. Once amplified, the DNA produced by
PCR can be used in many different laboratory procedures. For example,
most mapping techniques in the Human Genome Project (HGP) relied on
PCR. PCR is also valuable in a number of laboratory and clinical
techniques, including DNA fingerprinting, detection of bacteria or viruses
(particularly AIDS), and diagnosis of genetic disorders.
To amplify a segment of DNA using PCR, the sample is first heated so the
DNA denatures (separates into two pieces of single-stranded DNA). Then,
an enzyme called "taq polymerase" synthesizes two new strands of DNA,
using the original strands as templates. This process results in the
replication of the original DNA, with each of the new molecules containing
one old and one new strand of DNA. Then each of these strands can be
used to create two new copies and so on. The cycle of denaturing and
synthesizing new DNA is repeated as many as 30 or 40 times, leading to
more than one billion exact copies of the original DNA segment. The entire
process of PCR is automated and can be completed in just a few hours. It
is directed by a machine called a thermocycler, which is programmed to
alter the temperature of the reaction every few minutes to allow DNA
denaturing and synthesis. (Polymerase Chain Reaction (PCR), 2015)

The Process of DNA Fingerprinting

The procedure for creating a DNA fingerprint consists of first obtaining a
sample of cells, such as skin, hair, or blood cells, which contain DNA. The

Anuththara Gunatilaka - Biology

DNA is extracted from the cells and purified. Next the restriction enzymes
are used to cut the DNA into smaller pieces. Restriction enzymes work by
cutting the DNA at a specific sequence, which produces either blunt ends
or sticky ends, and results in many fragments of different lengths. These
fragments are called restriction fragments length polymorphisms, or
RFLPs.
These RFLPs are then put into an agarose gel. Using gel electrophoresis,
the fragments are sorted according to size. When the current of the
electric field is turned on, the negative RFLPs will start to move across the
gel towards the positive end. The smaller fragments move farther across
the gel than the larger ones. Also, alkali is responsible for causing the
hydrogen bonds to break, and the DNA to become single-stranded. When
the DNA becomes single-stranded, it causes nucleotides to become free,
and they will later be used to pair up with probes. The gel is then covered
by a piece of nylon and thin paper towels, which are used to absorb
moisture from the gel. The DNA fragments get gently transferred from the
gel to the surface of the nylon. This process is called blotting.
Finally, radioactive probes get washed over the nylon surface. These
probes will join to any DNA fragments that share the same
composition. The final step to making a genetic fingerprint is to place a
photographic film on top of the nylon surface. The probes leave marks on
the film wherever they attached to the RFLPs. Dark bands will then show
up when the film is developed, which marks the length of the RFLPs that
were hybridized. Researchers are then able to read the fingerprint and
match it to others. They do this by placing the X-ray on a light
background, and (for example) comparing the RFLP lengths in the DNA
from the crime scene, to the DNA of the suspect. (Britannica, 2014)

Anuththara Gunatilaka - Biology

Anuththara Gunatilaka - Biology

Application of DNA Fingerprinting

Criminal investigations:
In criminal investigations, the DNA fingerprint of a suspect's blood or other
body material is compared to that of the evidence from the crime scene to
see how closely they match. The Internet and computers allow law
enforcement officials to place prints in a massive database run by the
Federal Bureau of Investigation (FBI) that can be accessed from anywhere
in the world. The technological advances in fingerprinting have also had a
major impact on identifying prints at crime scenes as well, not just in
recording them from a person. As fingerprints are unique to each
individual, they serve as a highly accurate way for law enforcement
agencies to identify a suspect, as well as potentially prove their guilt or
innocence. (University, n.d.)
Paternity tests:
DNA fingerprinting can be used to establish if someone is the biological
parent of a child, where the DNA fingerprinting of a childs body material
is compared to that of his/her father in order to identify how closely they
match. This test also may be used to identify the parents of babies who
were switched at birth. (BBC, 2013)

Bibliography
(UOA), U. o. (2004, July). VNTR: hypervariable regions. Retrieved from The
Biology Project:
http://www.biology.arizona.edu/human_bio/problem_sets/dna_forensics_1/0
5t.html
BBC. (2013, February 1). How does DNA testing work? Retrieved from BBC
Science: http://www.bbc.co.uk/science/0/20205874
Britannica, T. E. (2014, December 12). DNA fingerprinting. Retrieved from
Encyclopdia Britannica: http://www.britannica.com/science/DNAfingerprinting
Polymerase Chain Reaction (PCR). (2015, June 16). Retrieved from National
Human Genome Research Institute: https://www.genome.gov/10000207
University, P. S. (n.d.). How Fingerprinting Improves Criminal Investigations.
Retrieved from CRIMINOLOGY AND CRIMINAL JUSTICE:
http://online.ccj.pdx.edu/resources/news-articles/how-fingerprintingimproves-criminal-investigations/