Nucleotide sequencing denaturation/annealing steps

Figure 5-47 Amplification of DNA using the polymerase chain reaction (PCR). Double-stranded DNA is denatured by heating to 90-99° C (step a) and oligonucleotide primers complementary to short 12-18 nucleotide sequences at the two ends of the piece of DNA to be amplified are annealed to the separated strands by cooling to 40 - 75° C (step b). The two DNA strands serve as templates for synthesis of new complementary strands using a heat-stable DNA polymerase and a mixture of the four nucleotide triphosphates. Nucleotide units are added to the 3 ends of the primers, with the new chains growing in the 5 3 ...

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