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Improve Amplification Specificity with Hot Start PCR Enzyme

ling and extension at non-restrictive temperatures produces undesirable products that are amplified throughout the remaining PCR cycles. Misprimed products and artifacts such as primer-dimers can impair gel analysis, quantitation, and sequencing of specific PCR products. Moreover, in the amplification of misprimed products, dNTPs and primers are diverted away from the synthesis of specific products, reducing overall yields.

Stratagenes SureStart Taq DNA polymerase improves amplification reactions by reducing background and increasing yield of desired product.

SureStart Taq DNA polymerase consists of a reversibly inactivated version of Stratagenes Taq2000 DNA polymerase, a highly purified recombinant Taq DNA polymerase preparation. SureStart Taq polymerase remains inactive until stringent denaturation temperatures are reached. It is activated by adding a heat step (9 to 12 minutes at 92 to 95C) to the beginning of thermal cycling programs (pre-PCR heat-activation method). Alternatively, the pre-PCR heat-activation step can be omitted to permit enzyme activation during temperature cycling. When the PCR heat-activation method is employed, it is generally necessary to add additional cycles to existing cycling programs to achieve optimal product yield.

SureStart Taq DNA polymerase provides superior or equivalent improvements in specificity and product yield compared to other vendors hot start Taq preparations. In the amplification system shown in the figure, Taq DNA polymerase synthesizes multiple extraneous bands in addition to the desired 105 bp fragment (lane 2). In contrast, background is eliminated in reactions employing SureStart Taq DNA polymerase (lane 1). SureStart Taq DNA polymerase provided improved specificity and higher product yield compared to other vendors antibody-based (lane 3) and modified (lane 4) Taq hot start formulations. Moreover, using SureStart Taq DN
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