Q.

Which of the following is a correct sequence of steps in a PCR (Polymerase Chain Reaction) ?

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a

Denaturation, annealing, extension

b

Annealing, denaturation, extension

c

Extension, denaturation, annealing

d

Denaturation, extension, annealing

answer is B.

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Detailed Solution

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The sequence of steps in polymerase chain reaction is denaturation, annealing and extension. The denaturation step generates templates by converting a dsDNA into ssDNA at high temperature (94°C). Annealing involves addition of primer and its hydrogen bonding with the template DNA at 72°C. Extension involves addition of dNMPs to the primer by polymerisation reaction carried out by heat stable DNA polymerase (72°C). When these three steps are repeated many times, the desired segment of DNA can be amplified approximately up to 1 billion copies.

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