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Greater Amplification Specificity with New Hot Start PCR Enzyme

he target from 500 and 5000 copies of HIV-1 template, nonspecific products predominated in reactions using 5, 50, and 500 copies (Figure 2). In contrast, SureStart Taq DNA polymerase produced specific amplification product from as few as five copies of DNA template, and no background was visible (Figure 2).

In Figure 3, a 970-bp fragment was amplified from 101 to 105 copies of lambda DNA in the absence or presence of 0.5 mg of denatured genomic DNA. In a complex DNA sample, multiple nonspecific products were generated, and specific product yield was reduced (Figure 3A). When compared to Taq, SureStart Taq DNA polymerase successfully amplified the desired product from as few as 101 copies of lambda DNA, even in the presence of high concentrations of extraneous denatured DNA. Moreover, SureStart Taq DNA polymerase generated far fewer nonspecific products compared to Taq DNA polymerase. Comparisons with other vendors hot start enzymes showed similar improvements in specific product yield (Figure 3B, Lanes 1 to 3) but high background was evident in two of the three hot start preparations tested.

Conclusions

Use Stratagenes SureStart Taq DNA polymerase to achieve a simple and effective method for hot start. SureStart Taq DNA polymerase improves the sensitivity of amplification reactions by reducing background and increasing yield of desired product. Compared to other vendors hot start Taq preparations, SureStart Taq provides comparable or superior detection of low-copy-number targets. SureStart Taq is inactive at ambient temp
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