PCR

The polymerase chain reaction (PCR) is a method to rapidly amplify sequences of DNA. During a typical PCR, template DNA (containing the region of interest) is mixed with deoxynucleotides (dNTPs), a DNA polymerase and primers. Primers are short segments of complimentary DNA that base-pair with the template DNA upstream of the region of interest and serve as recruitment sites for the polymerase. PCR involves a series of temperature cycles that, although once conducted by moving tubes through various water baths, is now controlled automatically by the use of thermal cyclers, or thermocyclers. Thermocyclers provide tight control over both the reaction temperature and the duration of each temperature step, ensuring efficient amplification. During a typical PCR, cycles of denaturation, annealing and extension are repeated to achieve exponential amplification of the target sequence. Denaturation consists of heating the samples up to a high temperature (typically 94-98°C) to cause denaturation of the template DNA, disrupting the hydrogen bonds and base stacking interactions that hold the DNA strands together. 

PCR includes these areas of focus:

Routine PCR
High-Fidelity PCR

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