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Greater Amplification Specificity with New Hot Start PCR Enzyme

g ramp-up to initial denaturation temperatures. In comparison, commercially available preparations of modified Taq remain inactive until higher temperatures are reached (>4 minutes at 95C 10). As with other hot start procedures, the use of reversibly inactivated DNA polymerases allows for PCR setup at room temperature, while providing reduced background and enhanced amplification of specific product. Moreover, hot start PCR enzymes can be introduced into existing amplification systems with little or no modification of PCR protocols, and their use avoids awkward manipulations associated with wax barrier or manual methods.

Eliminate Nonspecific PCR Products

SureStart Taq DNA polymerase is a chemically inactivated version of Stratagenes Taq2000 DNA polymerase1, a highly purified recombinant Taq DNA polymerase preparation. SureStart Taq DNA polymerase is provided in an inactive state and remains completely inactive until stringent denaturation temperatures are reached. The enzymes accompanying 10X reaction buffer has been formulated to provide flexible and efficient enzyme activation, resulting in optimal specificity and product yield. SureStart Taq DNA polymerase can be partially or completely activated in a pre-PCR heat step (9 to 12 minutes at 92 to 95C Pre-PCR Heat Activation Method) or it can be activated slowly during the denaturation steps of thermal cycling (PCR Heat Activation Method).

In Figure 1, results from the Pre-PCR and PCR Heat Activation Methods are compared. In this example, a 105-bp fragment of the glucocerebrosidase gene was amplified from human genomic DNA using Taq (Figu
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