rBst DNA Polymerase

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Last modified: 2008-07-15 14:57:15

rBst DNA Polymerase, produced from an over expressing recombinant clone in E. coli, is the product of the DNA pol I gene of the thermophilic bacterium Bacillus stearothermophilus (Bst). The enzyme has optimal activity at 65°C and at elevated temperatures it can synthesise DNA in regions of template secondary structure or high GC content where standard DNA polymerases may fail. These characteristics make rBst DNA Polymerase useful when replicating difficult templates. The enzyme has 5'Æ3' exonuclease activity. Based on work at Epicentre®, rBst DNA Polymerase is also active as a thermostable RNA-dependent DNA polymerase (reverse transcriptase).

Unit Definition:

One unit of rBst DNA Polymerase converts 10nmoles of deoxyribonucleotides into acid-insoluble material in 30 minutes at 65°C in 25mM TAPS, pH 9.3 at 25°C, 50mM KCl, 2.0mM MgCl2, 0.2mM of each dNTP, and 10µg activated calf thymus DNA.

Storage Buffer:

50% glycerol containing 50mM Tris-HCl, pH 7.5, 100mM NaCl, 0.1mM EDTA, 1mM DTT, and 0.1% Triton® X-100.

Quality Control:

rBst DNA Polymerase is tested for activity using various DNA templates and is free of detectable RNase and endonuclease activities.

Last modified: 2008-05-14 16:46:46

Applications

• First strand cDNA synthesis from primed RNA templates
• Production of templates for use in RT-PCR amplification reactions

Last modified: 2008-05-14 16:46:46

Protocols for: rBST DNA Polymerase

(catalogue number BH1100 / BH1500 / BH101K)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Stenesh, J. and Roe, B.A. (1972) Biochim. Biophys. Acta 272, 156.
2. Ye, S.Y. and Hong, G.F. (1987) Scientia Sinica 30, 503.

Last modified: 2008-05-14 16:46:46