PCR thermal cycle 2026-10-14
A PCR thermal cycle separates three jobs. During PCR denaturation, heating separates the paired DNA strands without intentionally cutting their phosphodiester bonds. During PCR annealing, cooling permits base pairing between PCR primers and templates. During PCR extension, the chosen interval supports DNA polymerase synthesis from the paired ends. Each new nucleotide is supplied as a deoxynucleoside triphosphate; its incorporation lengthens the strand. The DNA polymerase is a catalyst, while these nucleotides are consumed substrates.[ref]
Taq polymerase withstands repeated high-temperature strand separation better than a heat-sensitive DNA polymerase. Its extension activity still depends on temperature, buffer solution and magnesium ions. Heat resistance does not mean constant activity at every stage: the separation temperature serves the DNA, whereas the extension temperature serves the copying enzyme. As one institutional example, Addgene's Basic PCR Program specifies separation, primer binding and extension. These are example settings, not universal properties of all PCR primers or DNA polymerases.[ref]