Extremely thermostable DNA polymerase, enabling efficient high fidelity PCR of genomic targets up to 12 kb and episomal targets up to 20 kb.
Description:
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Hybrid is a genetically engineered thermophilic DNA polymerase.
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Ultrapure recombinant enzyme.
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The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5'->3' direction in the presence of magnesium ions.
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The enzyme exhibits the 3'->5' proofreading activity, resulting in over 10-fold higher PCR fidelity than possible with Taq DNA Polymerases.
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The enzyme generates blunt ends.
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Enhanced polymerase processivity allows to use shorter extension times.
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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).
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Due to the genetic modification of the polymerase, the optimal reaction conditions (especially annealing temperatures) differ from standard PCR protocols.
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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.