Question

Explain the steps involved in PCR amplification method.

Hint:

Remember the different temperatures used in different steps of PCR, the functions of primers and DNA polymerase, and how the process leads to amplification of a sequence.

Answer 1

Denaturation: DNA is heated to 94-98°C, separating it into single strands to prepare for amplification. Annealing: Temperature is reduced to 50-65°C, allowing primers to bind to specific sites on the single-stranded DNA templates. Extension: At 72°C, optimal for Taq DNA polymerase, primers are extended using dNTPs, replicating the target DNA sequence. Repeat Cycles: This cycle is repeated many times, leading to exponential amplification of the target DNA, generating millions of copies.