Polymerase Chain Reaction: Basic Principles and Routine Practice

The polymerase chain reaction (PCR) is a primer-mediated enzymatic amplification of specifically cloned or genomic DNA sequences (1 ). This PCR process, invented by Kary Mullis over 10 years ago, has been automated for routine use in laboratories worldwide. The template DNA contains the target sequence, which may be tens or tens of thousands of nucleotides in length. A thermostable DNA polymerase, Taq DNA polymerase, catalyzes the buffered reaction in which an excess of an oligonucleotide primer pair and four deoxynucleoside triphosphates (dNTPs) are used to make millions of copies of the target sequence. Although the purpose of the PCR process is to amplify template DNA, a reverse transcription step allows the starting point to be RNA (2 –5 ).