What is the number of cycles of three step 'Denaturation - Annealing - Extension' needed in Polymerase Chain Reaction (PCR)?

Expert Solution

Theoritically, as DNA copies are doubled at each cycle completion, after n PCR cycles 2ⁿ copies are generated from an original DNA copy. However, PCR efficiency varies. The number of cycles needed for a particular purpose depends on the number of starting DNA template copies, extension and amplification efficency of the reaction and the desired final DNA yield.

About 30 3-step PCR cycles are sufficient if starting DNA is about 10⁵ template sequence copies and (Taq) DNA polymerase efficiency is near 0.7. The yield of the final product thus generated is sufficient to visualize on the gel or analysis by DNA sequencing. With low starting DNA amounts, upto 40 PCR cycles may be carried out. But high number of cycles should be avoided as factors like non-specific binding, byproduct accumulation, reduced efficiency come into play.

About 30 3-step PCR cycles are sufficient if starting DNA is about 10⁵ template sequence copies and (Taq) DNA polymerase efficiency is near 0.7.

Shams answered 2 years ago

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