Pfu DNA Polymerase（Wth HighPure dNTP Mix)

Catalog# BPC1023-2

Lot # Check on the product label

Size: 3000 U

Storage: Store at -20°C for one year. 

Concentration: 5 U/µl

Description: Pfu DNA Polymerase is isolated from E.coli strain that contains Pyrococcud fureosis DNA Polymerase gene.  Pfu DNA Polymerase possesses a proofreading 3'-5' exonuclease activity. Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis.

Kit Components

Activity Definition: One unit of EasyPfu DNA Polymerase incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material at 74°C for 30 min.

Storage Buffer: 50 mM Tris-HCl (pH 8.0), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, stabilizers, and 50% (v/v) glycerol.

A10X Pfu Buffer with 20 mM MgSO₄: 200 mM Tris-HCl (pH 8.8), 100 mM (NH4)₂SO₄, 100 mM KCl, 1% (v/v) Triton X-100, 1mg/ml BSA and 20 mM MgSO₄.

Precatious:

1. Because 3'-5' exonuclease activity, Pfu DNA polymerase extension rate is lower than Taq DNA polymerase. Therefore, during the extension step, please set extension time according to the product length.

2. High primer purity and concentration (about 0.2-0.5μM) required. The 3'-5' exonuclease activity may degrade primers, especially, when the reaction mixture lack of dNTP. To overcome the degradation, add the Pfu last into the reaction mixture and carry PCR reaction at once.

3. The stability of Pfu DNA Polymerase is higher than Taq DNA Polymerase. For the CG rich template, the denaturation temperature can increase to 98°C without affecting the activity of Pfu DNA polymerase.

4. Pfu DNA polymerase produces blunt-ended amplification products. The products can be cloned by blunt-ended vector not by T/A vector.

PCR mixture set up: (for 50 μl reaction volume)