Hybrid DNA Polymerase (Extremely thermostable DNA polymerase, enabling efficient high fidelity PCR of genomic targets up to 12 kb and episomal targets up to 20 kb)

Extremely thermostable DNA polymerase, enabling efficient high fidelity PCR of genomic targets up to 12 kb and episomal targets up to 20 kb.

Description:

• Hybrid is a genetically engineered thermophilic DNA polymerase.
• Ultrapure recombinant enzyme.
• The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5'->3' direction in the presence of magnesium ions.
• The enzyme exhibits the 3'->5' proofreading activity, resulting in over 10-fold higher PCR fidelity than possible with Taq DNA Polymerases.
• The enzyme generates blunt ends.
• Enhanced polymerase processivity allows to use shorter extension times.
• The modification of Hybrid DNA Polymerase enhances target length capability of PfuPlus! DNA Polymerase with regard to genomic targets (up to 12 kb from human genomic DNA).
• Due to the genetic modification of the polymerase, the optimal reaction conditions (especially annealing temperatures) differ from standard PCR protocols.
• Hybrid DNA Polymerase is recommended for general use in PCR, use in high-fidelity PCR, PCR of GC-rich sequences or problematic secondary structures and cloning of blunt-ended PCR products.

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 min at 74°C. The reaction conditions are: 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 and 0.1 mg/ml BSA in a final volume of 50 μl.

Storage Conditions: Store at –20°C.

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.

10 x Reaction Buffer:

10 x Hybrid Buffer: the buffer contains 15 mM MgCl₂.

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.