Hybrid DNA Polymerase

Description

• Ultrapure recombinant enzyme
• Catalyzes the polymerization of nucleotides into duplex DNA in the 5'->3' direction in the presence of magnesium ions
• Exhibits 3'->5' proofreading activity, resulting in an over 10-fold higher PCR fidelity than standard Taq DNA Polymerases
• Generates blunt ends
• Enhanced polymerase processivity allows shorter extension times

Applications

• For use in standard and High-fidelity PCR
• PCR of GC-rich sequences
• Problematic secondary structures
• Cloning of blunt-ended PCR products

Note

Due to the genetic modification of Hybrid DNA Polymerase, the optimal reaction conditions, including annealing temperatures, differ from standard PCR protocols 

Unit Definition

One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 minutes at 74°C

Reaction Conditions

50 mM Tris-HCl (pH 9.0 at 25°C)
50 mM NaCl
5 mM MgCl₂
200 μM each of dATP, dCTP, dGTP, dTTP (a mix of unlabeled and [3H]dTTP)
10 μg activated calf thymus DNA
0.1 mg/ml BSA in a final volume of 50 μl

Storage Buffer

20 mM Tris-HCl (pH 8.0 at 22°C)
100 mM KCl
0.5% Tween™20
0.5% Igepal CA-630
0.1 mM EDTA
1 mM dithiothreitol
50% glycerol and stabilizers

10X Hybrid DNA Polymerase Reaction Buffer

Contains 15 mM MgCl₂

PCR amplification using CHIMERx Hybrid DNA Polymerase

Lane M: molecular size marker - Perfect 1 kb DNA Ladder. Lanes 1.1 to 12 kb: PCR amplification reactions, using 1 unit of Hybrid DNA Polymerase and 100-500 ng of human genomic DNA as a template for 35 cycles in 50 μl reaction volume.

Quality Control

All preparations are assayed for contaminating 3'-exonuclease, and nonspecific single- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.

Storage Conditions

Store at -20ºC
Product shipped on dry ice