The Design and Optimization of the PCR

In the few years since its introduction,1,2,3 the polymerase chain reaction has already become a widespread research technique. Like the PCR itself, the numbers of its practitioners have been accumulating exponentially and will probably continue to do so in the near future as the method finds wider applications in fields other than molecular biology. This popularity of the PCR is primarily due to its apparent simplicity and high probability of success. Reduced to its most basic terms, PCR merely involves combining a DNA sample with oligonucleotide primers, deoxynucleotide triphosphates, and the thermostable Taq DNA polymerase in a suitable buffer, then repetitively heating and cooling the mixture for several hours until the desired amount of amplification is achieved.

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[4]  H A Erlich,et al.  Direct cloning and sequence analysis of enzymatically amplified genomic sequences. , 1986, Science.

[5]  I. Tinoco,et al.  Comparison between DNA melting thermodynamics and DNA polymerase fidelity. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[6]  K. Mullis,et al.  Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. , 1986, Cold Spring Harbor symposia on quantitative biology.