Bst DNA Polymerase (Large Fragment, exo -)
Source: Bacillus stearothermophilus
Large exonuclease free fragment of thermophilic Bst DNA Polymerase with strand displacement activity.
Description:
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Bst DNA Polymerase is a moderately thermostable enzyme from Bacillus stearothermophilus.
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Ultrapure, recombinant protein.
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The enzyme replicates DNA optimally at 65°C.
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Catalyzes the polymerization of nucleotides into duplex DNA in the 5´->3´ direction in the presence of magnesium ions.
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Lacks the 5´->3´ exonuclease activity, while retaining the polymerase activity (1).
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Broad activity range; can replace mezophilic polymerases as well as synthesize DNA at high temperatures. Thus it is suitable for amplification of difficult DNA templates, including repetitive sequences, GC-rich regions and problematic secondary structures (2,3).
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Can be heat inactivated at temperatures above 80°C.
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Active over wide range of reaction buffer conditions and magnesium ions concentrations.
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Used in isothermal DNA sequencing at elevated temperatures.
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Ideal for DNA synthesis reactions requiring strand displacement.
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Exhibits thermophilic reverse transcriptase activity.
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Used in isothermal nucleic acids amplification.
Unit Definition: One unit is the amount of enzyme required to incorporate 10 nmoles of total deoxyribonucleotide into acid-insoluble form in 30 min at 60°C.
Storage Conditions: Store at –20°C.
1 x Reaction Buffer: 50 mM Tris-HCl, (pH 8.9 w 20oC), 10 mM (NH4)2SO4, 10 mM KCl, 2 mM MgSO4, 0.1% Triton™X-100.
Storage Buffer: 20 mM potassium phosphate (pH 6.8), 1 mM dithiothreitol and 50% (v/v) glycerol.
Quality Control: All preparations are assayed for contaminating endonuclease, exonuclease and single- and double-stranded DNase activities.
References:
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Stenesh, J. and Roe, B.A. (1972) Biochim. Biophys. Acta. 272, 156-166.
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Hugh, G. and Griffin, M. (1994) PCR Technology, p.p.228-229.
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McClary, J. et al. (1991) J. DNA Sequencing and Mapping, p.p.173-180.