DNA Fingerprinting
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It
is way to identify a species or particular trade.
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A
technology used to by scientist to distinguished between individual to same
species using samples of the DNA.
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Discovered
by Allex Jeffryesin 1986.
Steps of
DNA-Fingerprinting
There are eight
steps of DNA-Fingerprinting.
1] Isolation
of DNA –
DNA must be
recovered from cells, only a small amount of blood ski needed to isolate DNA.
2] digestion
of DNA by restriction endonuclease -
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The
DNA is cut from fragments by using restriction enzyme.
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Each
restriction enzyme cut`s DNA at a specific base sequence.
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These
sections of that DNA cut out are called restriction enzyme.
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Reaction
mixture are incubated at night 370c.
3] Electrophoretic
separation of different fragment –
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Fragment
are separated on the basis of size using a process is called gel
electrophoresis.
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DNA
fragment are injected in small wells on the apply electric current along the
gel
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DNA
is negative charge then this DNA move to words in positive direction end of the
gel.
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Small
DNA fragment fast move than the large fragment.
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DNA
is separated on the basis of size.
4] Transfer
DNA on nitrocellulose –
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The
DNA fragments are transfer in nylon sheet by placing the sheets on gel and
socking them overnight by the process is called southern blotting.
5] Probe labelling –
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Adding
probe/colored probe in nylon sheet which is complementary in DNA fragment.
6] Hybridization –
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Labelled
probe DNA should be hybridized with the complementary sequence located on nylon
membrane for the detection of position of factor membrane washed to remove
non-specific binding and the cleaning of the background.
7] Autoradiography –
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To
detect the sequence in genome bound with hybridized, radioactive probe on
membrane.
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The
X-ray film after the developed shows multiple number of bonds that look like
bar codes and known as DNA fragments.
8] Interpretation of bond pattern –
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Analysis
bond pattern of different individual comparison of position bond.
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Computer
software also available for the analysis of DNA fingerprints.
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