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The page below is a sample from the LabCE course Real-Time PCR. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Steps of PCR - Denaturation

Denaturation is the process that causes the paired strands of DNA to separate into individual single strands.
The reaction mixture is heated to approximately 95°C for 20-30 seconds in order to promote DNA denaturation. At this point, the hydrogen bonds within the DNA break, resulting in single-stranded DNA molecules. The Taq polymerase is still able to survive at this temperature, but will not activate.