Fundamentals of the Polymerase Chain Reaction and

Fundamentals of the Polymerase Chain Reaction and Separation of the Reaction Products 

Since its introduction, the PCR technique (Saiki et al., 1985 Mullis and Faloona, 1987) has been used extensively in molecular biology to amplify specific 

DNA sequences that are present in trace quantities. While the design seems simple—repeated cycles of denaturation, primer binding, and DNA synthesis—this powerful technique can result in lf -fold amplification from a single copy of target DNA. 

Essentially, all forms of DNA (and RNA) can be amplified by PCR. The amount of DNA template required for amplification depends on the complexity of the genome. Typically, 0.1 to 1 mg of mammalian genomic DNA is required for PCR, while only picogram to nanogram quantities are utilized for bacteria (Sambrook et al., 1989). 

DNA polymerase is required for synthesis. There are several different polymerases available, each with its 6Wn characteristics that will affect its efficacy in the PCR. Since many labs have studied the properties of the different polymerase in current use, much is known about their efficiencies in PCR and rate of base misincorporation (Table 7.1). The thermostable polymerases (Tag, Vent) are preferred in PCR over the heat-labile enzymes 

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