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Identification of an individual rely on hypervariable regions which are segments of individuals

which vary can greatly between two individuals


Hypervariable regions which are used for DNA profiling include:
STR's (short tandem repeats)/microsatellites - Short tandem repeats are short sequences (2-5
base pairs) that repeat within a chromosome over and over sequentially
Number of repeats at a STR varies between individuals and each variations is 'a distinct allele',
hundreds are scattered throughout human chromosomes. One may be homozygous or
heterozygous for number of short tandem repeats at a particular locus (5 repeats to 7), however
many different combinations of alleles arise within the gene pool of a population (e.g. individual I
14,8 and individual II 15,10 for particular locus)
- STR's are very sensitive, based on alleles whose fragments may vary by only one base pair, is
carried out by one single-locus probe and carried out in a much shorter time
mtDNA hypervariable regions mtDNA identification is less precise/less unique as mtDNA is
carried through person's matrilineal line hence many individuals can have same mtDNA
What are probes?
Probes are radioactively labeled known short strands of DNA which bind to complementary
sequences when introduced to a solution of DNA. They are used to locate certain fragments
within a DNA sequence
The process of Polymerase chain reaction (PCR) allows for the production of mass quan
tities of genetic material from a minute starting quantity. PCR can be used for medical
applications (testing for genetic mutations) and in forensics.
The process of PCR revolves around the repetition of 3 main parts;
- Denaturation
- Annealing and
- Elongation
During the denaturation stage; the DNA is heated to temperatures of around 90C in order to
break hydrogen bonds between the adjacent DNA strands
During the annealing stage; Primer's (composed of short synthetic DNA strands) with nucleic
acid sequences complementary to a portion of the DNA strand hybridize to their corresponding
segments of DNA. Temperatures of around 55C are used during this stage
During the elongation stage; DNA polymerase (TAQ polymerase) binds to a forward/reverse
primer and begins to synthesis a new strand of DNA using nucleotides found in solution, with a
base sequence that is complementary to the template DNA strand. Temperature of 65C are
used for optimal enzyme activity.
These three stages are repeated consecutively with the quantity of genetic material increasing
in accordance with the exponential function 2^n, where n= number of times process is
completed
Electrophoresis is used to obtain a genetic profile and has many applications such as in
paternity testing, and forensics
Electrophoresis separates DNA strands on the basis of length(size/mass)
Electrophoresis is usually performed after PCR has been completed

Genetic material is first subject to restriction enzymes which cuts the DNA sequence in
accordance to their recognition sequences [restriction enzymes may be also used to introduce a
DNA fragment into a bacterial plasmid as plasmid will be cut over the same recognition
sequence - blunt ends/sticky ends]
DNA samples from the subjects alongside a control (for size measurement) are suspended
inside holes (made by a comb) within an agarose gel plate
A negative terminal is placed adjacent to a positive terminal on either side of the agarose gel
plate. The DNA samples are located at the negative terminal
Current is switched on, and segments of DNA from each subject proceed to move towards the
opposing pole at a pace in accordance with their size and charge
After a suitable period of time, current is switched off and a DNA profile from each subject can
be obtained, and matched if required