Manufactur standard Qpcr Taqman - 2×Pfu PCR Mix – TIANGEN

Activity Definition

The activity of 1 unit (U) Pfu DNA Polymerase is defined as the amount of enzyme required to incorporate 10 nmol deoxynucleotides into acid-insoluble substances at 74°C within 30 min using activated salmon sperm DNA as a template/primer.

Quality Control

The purity by SDS-PAGE detection is more than 99%; No activity of exogenous nuclease is detected; Single-copy gene in human genome could be amplified effectively; No significant activity change when stored at room temperature for one week.

Main Technical Parameters

It has 3′-5′ exonuclease activity and no 5′-3′ exonuclease activity. The extension speed of DNA amplification is lower than that of Taq polymerase, and generally the extension speed of Pfu enzyme is 0.5-1 kb per minute. The thermal stability of Pfu is better than Taq. For templates with high GC content, the denaturation temperature can be increased to 98°C, which has no effect on the activity of Pfu polymerase. The PCR product is blunt-ended, which can be added with 3’-dA overhangs before ligated with TA vector or cloned with blunt-ended vector

Applications

It can be used for high fidelity amplification of DNA, such as gene expression cloning, site-directed mutation, single nucleotide polymorphism (SNP) analysis and end repairing.

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