PCR Performance Comparisons Between pfuturbo and Taq DNA,,,Polymerases ( -6 and six-fold lowerthan that of T...)

PCR Performance Comparisons Between pfuturbo and Taq DNA,,,Polymerases

-6 and six-fold lower than that of Taq DNA polymerase.³ PfuTurbo DNA polymerase exhibits the same high-fidelity benefits as Pfu DNA polymerase.¹

Conclusions

In side-by-side comparisons, we determined that PfuTurbo DNA polymerase amplified longer complex targets (>3 kb) in higher yield than Taq DNA polymerase. High-fidelity amplification of genomic targets up to 10 kb, and cloned targets up to 15 kb, were previously achieved with PfuTurbo DNA polymerase.¹ Although PfuTurbo and Taq DNA polymerases can amplify relatively easy targets (0 to 2 kb) with comparable yields, higher replication fidelity makes PfuTurbo DNA polymerase the better choice for amplifying DNA targets to be cloned, expressed, and/or sequenced. Thus, adding a novel PCR-enhancing factor makes PfuTurbo DNA polymerase superior to Taq DNA polymerase for both routine and high-performance PCR applications.

Methods

PCR amplifications were conducted with PfuTurbo DNA polymerase or Taq DNA polymerase in their recommended buffers, using 200 M each dNTP, 100 to 300 ng of genomic DNA or 15 ng of lambda phage DNA, and 100 ng of each oligonucleotide primer per 50 l reaction. PCR reactions were conducted in Stratagenes RoboCycler Gradient 96 temperature cycler fitted with a Hot Top assembly, using 200 l thin-walled PCR tubes. The temperature cycling parameters for all targets incorporated the following: one cycle at 95C for one minute, followed by 30 cycles at 95C for one minute (denaturation); 58 to 64C for one minute (annealing); 72C for one minute per kb of target amplified (extension); and one final extension cycle of 72C for 10 minutes. The amplified reaction products were electrophoresed on a 1% agarose/1X TBE gel, stained with ethidium bromide, and imaged using the Eagle EyeII Still Video System. Lanes labe
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( -6 and six-fold lowerthan that of T...)PCR Performance Comparisons Between pfuturbo and Taq DNA,,,Polymerases

Conclusions

Methods