During this stage the reaction is cooled to 50-65⁰C.This usually takes between 15-30 seconds.It is important that the temperature is maintained at this stage for long enough to ensure that the DNA strands have separated completely.This results in two single strands of DNA, which will act as templates for the production of the new strands of DNA.The high temperature causes the hydrogen bonds between the bases in two strands of template DNA to break and the two strands to separate.During this stage the cocktail containing the template DNA and all the other core ingredients is heated to 94-95⁰C.Image credit: Genome Research Limited What happens at each stage of PCR? Denaturing stage Illustration showing the main steps in the polymerase chain reaction (PCR). After PCR has been completed, a method called electrophoresis can be used to check the quantity and size of the DNA fragments produced.
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