Hiphi¢â DNA Polymerase
Description
Mbiotech HiPhi¢â DNA Polymerase is an extremely thermostable proofreading DNA polymerase, suitable for applications requiring high temperature synthesis of DNA. DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5¡¯ to 3¡¯ direction in the presence of Mg2+. It exhibits the 3¡¯ to 5¡¯ proofreading activity, resulting in over 10-fold higher fidelity than possible with Taq DNA Polymerase
Features
• Ultra pure recombinant protein.
• Recommended for use in high-fidelity amplification, amplification of GC-rich senquences or problematic secondary structures, primer extension reactions at elevated temperatures and cloning of blunt-ended amplification products.
• Recommended for use in high-fidelity amplification, amplification of GC-rich senquences or problematic secondary structures, primer extension reactions at elevated temperatures and cloning of blunt-ended amplification products.
Applications
• Routine PCR applications
• Primer extensions
• Primer extensions
10X PCR Buffer without MgCl2
500mM KCl, 100mM Tris-HCl (pH 9.1 at 20¡ÆC) and 0.1% Triton¢âX-100. The buffer is optimized for use with 0.1 - 0.2mM of each dNTP.
Storage Buffer
20mM Tris-HCl (pH 8.0 at 22¡ÆC), 100mM KCl, 0.5% Tween¢â 20, 0.5% Nonidet-P40, 0.1mM EDTA, 1mM DTT and 50% glycerol.
Storage and Stability
The Hiphi¢â DNA Polymerase is shipped on Dry/Blue Ice. All kit components should be stored at -20¡É upon receipt. Excessive freeze/thawing is not recommended. When stored under optimum conditions, the reagents are stable for a minimum of 6 months from date of purchase.
Quality Control
All preparations are assayed for contaminating endonuclease, exonuclease, and non-specific DNase activities. Functionally tested in DNA amplification.
Safety Precautions
Harmful if swallowed. Irritating to eyes, respiratory system and skin. Please refer to the material safety data sheet for further information.
Unit Definition
1u is defined as amount of enzyme that required to catalyze the incorporation of 10nmoles of dNTP into acid-insoluble material in 30 minutes at 74¡ÆC.
Figure
Amplification using Hiphi¢â DNA Polymerase
M1: 1 kb DNA Ladder
0.5 and 1.5: 0.5 kb ampification product generated using 0.2 mM dNTPs and 2.0u Pfu DNA Polymerase.
5 and 8: 5 kb and 8 kb amplification products generated using 0.25 mM dNTPs, 2.5u Pfu DNA Polymerase and 3% of formamide.
M2: Lambda / Hind III Marker
0.7% TAE agarose gel