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Hot Start PCR

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.
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).
Ease of JumpStart™ Versus Other Hot Start Methods

Product #

Description

Add to Cart

04743814001 Expand 20 kbPLUS PCR System, dNTPack
D5809 JumpStart AccuTaq LA DNA Polymerase Hot-start high fidelity Taq enzyme, 10X buffer included
D1313 JumpStart REDAccuTaq® LA DNA Polymerase Long and accurate hot-start Taq with inert dye, 10X buffer included
D8187 JumpStart REDTaq® DNA Polymerase Hot-start Taq enzyme with inert dye, 10X buffer included
P0982 JumpStart REDTaq® ReadyMix Reaction Mix for PCR
P1107 JumpStart REDTaq® ReadyMix Reaction Mix for High-throughput PCR of complex templates
A7721 JumpStart Taq Antibody Adds hot-start capabilities to any Taq DNA Polymerase
D9307 JumpStart Taq DNA Polymerase with MgCl2
D4184 JumpStart Taq DNA Polymerase without MgCl2
P2893 JumpStart Taq ReadyMix Complete optimized reagent for hot-start PCR at 2X concentration