... cycle, in order to continue amplification of the target sequence. Advances in this process include the use of thermostable DNA polymerases, which resist denaturation at high temperatures and so an initial aliquot if polymerase can last for the successive cycles needed in the PCR as well as the development of thermal cyclers or PCR machines, which rapidly change temperature as needed, in an automated programmable manner, resulting in modern PCR. The theory of PCR The DNA to be amplified, the template DNA, is denatured to separate each strand by heating at a temperature between 90-95ºC and this separation of the duplex allows the chosen oligonucleotide primers to anneal to their specific homologue within the template sequence. The primers should have a melting temperature (Tm) that allows annealing at temperatures of 55°-65°C and are designed to provide starting point for the synthesis of new DNA strands, complementary to the target DNA. If the ...

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