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Optimizing pfuturbo DNA Polymerase Amplification Reactions with,,,Perfect Match PCR Enhancer

ble single-enzyme formulations.1,2

Stratagenes PCR-enhancing Agents

Perfect Match PCR enhancer improves the yield and specificity of PCR amplification reactions carried out with Taq DNA polymerase.3,4,5 Perfect Match enhancer reduces or eliminates nonspecific amplification, possibly by destabilizing mismatched primer-template complexes that can lead to false priming and the generation of undesired PCR artifacts. While Perfect Match enhancer improved amplifications from both plasmid and genomic DNA templates, these optimal results depended on the ratio of Perfect Match enhancer to DNA template employed.4 Additional studies revealed that Perfect Match enhancer also improves amplifications of longer genomic DNA targets (2 to 5 kb) using Taq DNA polymerase; presumably by reducing secondary structure formation in the template strand, which impedes translocation of the DNA polymerase and limits amplification of longer targets.5

Perfect Match PCR enhancer and the Turbo additive in the PfuTurbo DNA polymerase formulation are distinct factors, which mediate PCR enhancement via different mechanisms. It is therefore likely that using Perfect Match enhancer could further improve the performance of PfuTurbo DNA polymerase, as has been demonstrated previously for Taq DNA polymerase PCR amplifications.3,4,5 Below we describe the effects of Perfect Match enhancer on amplifications of several difficult primer-template systems using PfuTurbo DNA polymerase.

Adding Perfect Match Enhancer to PCR Reactions

PCR amplifications were conducted with 5 U of PfuTurbo DNA polymerase or Taq2000 DNA in the enzymes recommended buffer, using 200 M each dNTP, 100 ng of genomic DNA, 100 ng of each oligonucleotide primer, and 1 U (1 l) of Perfect Match PCR enhancer per 50-l reaction. A 120-bp portion of the glucocerebrosidase gene was
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