Hot Start Polymerase Chain Reaction (PCR)
Michael R. Green, Joseph Sambrook
Abstract
Michael R. Green, Joseph Sambrook
Abstract
The purpose of hot start polymerase chain reaction (PCR) is to optimize the yield of the desired amplified product in PCRs and, simultaneously, to suppress nonspecific amplification and formation of primer dimers. This is achieved by withholding an essential component of the PCR-the DNA polymerase, or the primers, for example-until the reaction mixture has been heated to a temperature that inhibits hybridization of primers to one another or to nonspecific regions of the template.
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The purpose of hot start polymerase chain reaction (PCR) is to optimize the yield of the desired amplified product in PCRs and, simultaneously, to suppress nonspecific amplification and formation of primer dimers. This is achieved by withholding an essential component of the PCR-the DNA polymerase, or the primers, for example-until the reaction mixture has been heated to a temperature that inhibits hybridization of primers to one another or to nonspecific regions of the template.
Key concepts: Hot start PCR, Polymerase chain reaction optimization, Polymerase chain reaction, Primer (cosmetics), Inverse polymerase chain reaction, Polymerase, Molecular biology, Primer dimer