1.

Explain the principle of PCR (Polymerase Chain Reaction).

Answer»

PCR reaction is a DNA synthesis reaction that depends on the extension of primers annealed to opposite strands of a dsDNA template that has been denatured (melted apart) at temperatures near boiling. By repeating the melting, annealing and extension steps, several copies of the original template DNA can be generated.

The amount of starting material (target) needed is very small. 

Not necessary to isolate the desired sequence, because it will be defined by the primers that are used in the reaction. The primers are oligonucleotides complementary to different regions on the 2 strands of DNA template (flanking the region to be amplified). 

The primer acts as a starting point for DNA synthesis. The oligo is extended from its 3' end by DNA polymerase. 



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