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PCR principles

Using primers containing sequences that were not completely complementary to the template turned PCR into a tool for in vitro mutagenesis.
Replacing a single polymerase with a blend of a thermostable polymerase (Taq DNA Polymerase) and a proofreading polymerase (Pwo DNA Polymerase) made PCR an indispensable tool in the analysis and mapping of entire genomes by:
  • Extending the length of the sequence that could be amplified
  • Increasing the amount of PCR product
  • Providing higher fidelity during PCR
Using a Hot Start approach minimized the formation of primer-dimers during PCR.
Using short primers to produce a genomic fingerprint allowed analysis of organisms in which genomic sequences are largely unknown [e.g. Differ
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