Gene Amplification Through Polymerase Chain Reaction,DNA Segment,DNA Polymerase,Pyrococcus Furiosus,Thermococcus Litoralis

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	Gene Amplification Through Polymerase Chain Reaction - The polymerase chain reaction (PCR) technique, developed by Kary Mullis in 1985, is extremely powerful. It generates microgram (μg) quantities of DNA copies (upto billion copies) of the desired DNA (or RNA) segment, present even as a single copy in the initial preparation, in a matter of few hours. The PCR process has been completely automated and compact thermal cyclers are available in the market. The PCR utilizes the following: (1) a DNA preparation containing the desired segment to be amplified,
(2) two nucleotide primers (about 20 bases long) specific, i.e., complementary, to the two 3'-borders (the sequences present at the 3'-ends of the two strands) of the desired segment, (3) the four deoxynucleoside triphosphates, viz., TIP (thymidine triphosphate), dCTP (deoxycyctidine triphosphate), df\TP (deoxyadenosine triphosphate) and dGTP (deoxyguanosine triphosphate), and (a) a heat stable DNA polymerase, e.g. Taq (isolated from the bacterium Thermus acquaticus), Pfu (from Pyrococcus furiosus) and Vent (from Thermococcus litoralis) polymerases. Pfu and Vent polymerases are more efficient than the Taq polymerase.