Hot Start PCR

References
1. D’Aquila, R.T. et al. Maximizing sensitivity and specificity of PCR by pre-amplification heating. Nucleic Acids Res. 19, 3749 (1991).
2. Chou, Q., Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 20, 1717-1723 (1992).

In conventional PCR, the Taq DNA polymerase is active at room temperature and to a lesser degree, even on ice. In some instances, when all the reaction components are put together, nonspecific primer annealing can occur due to these low temperatures. This nonspecific annealed primer can then be extended by the Taq DNA polymerase, generating nonspecific products and lowering product yields.

Hot Start PCR significantly reduces nonspecific priming, the formation of primer dimers, and often, increases product yields. In Hot Start Long and Accurate PCR, the impact on yield can be dramatic. Classic methods, while effective, involve additional handling and increased risk of contamination. Sigma’s products for Hot Start PCR overcome all of these issues. JumpStart™ utilizes a neutralizing antibody to completely inactivate the Taq polymerase. The polymerase activity is completely restored during the first denaturation step of thermal cycling. This JumpStart is available for our standard Taq and REDTaq DNA™ polymerases as well as our Long and Accurate polymerase mixes. The neutralizing antibody is also available separately.