Hot Start PCR

Hot Start PCR is a more sensitive technique than standard PCR that allows amplification of low-abundance targets and single-copy genes while reducing PCR background problems.

Thermophilic DNA polymerases are unfortunately active at room temperature, which can result in amplification of unspecific targets due to random primer annealing events. Hot Start enzymes are completely inactive during room-temperature reaction setup and become active only after heating. Thus, misprimed amplification products and primer–dimer formation do not occur, diminishing PCR background.

Hot Start enzymes are typically created with one of the following techniques: chemically, by the addition of thermo-labile blocking groups, or with binding by one or more Taq-specific antibodies. Both of these techniques prevent enzyme activity until the blockage is disrupted at high temperature.

Usually hot-start modification does not affect PCR amplification: after thermal reactivation, hot start Taq DNA polymerases perform similarly to unmodified Taq with respect to processivity, fidelity and deoxyribonucleotidyl transferase activity, allowing TA cloning.

Hot start PCR master mixes containing all necessary PCR reagents are offered for simplified reaction setup. Green colored hot start PCR buffer is available for direct loading of PCR products onto gels.

 More information:

• Endpoint PCR Technical Guide
• Endpoint PCR Troubleshooting
• Endpoint PCR Technique Selection Guide

Hot Start PCR Product Selection Guide                                                          

NOTE: All above enzymes ensure the same PCR Fidelity like Taq DNA Polymerase, produce PCR products of up to 3 kb
with A overhangs suitable for TA cloning.