Details
Source
Bacillus stearothermophilus
Description
- Moderately thermostable enzyme from Bacillus stearothermophilus
- Bst DNA Polymerase, Large Fragment exhibits thermophilic reverse transcriptase activity
- Active over a wide range of reaction buffer conditions and magnesium ion concentrations
- Ultrapure recombinant protein
Applications
- Ideal for DNA synthesis reactions requiring strand displacement
- Used in isothermal nucleic acids amplification
- Used in isothermal DNA sequencing at elevated temperatures
- Ccatalyzes the polymerization of nucleotides into duplex DNA in the 5→3 direction in the presence of magnesium ions
- Lacks the 5→3 exonuclease activity, while retaining the polymerase activity (1)
- Broad activity range; can replace mesophilic polymerases as well as synthesize DNA at high temperatures, therefore suitable for amplification of difficult DNA templates, including repetitive sequences, GC-rich regions and problematic secondary structures (2, 3)
- Can be heat inactivated at temperatures above 80°C
- Replicates DNA optimally at 65°C
Unit Definition
One unit is the amount of enzyme required to incorporate 10 nmoles of total deoxyribonucleotide into acid-isoluble form in 30 minutes at 60°C.
Storage Buffer
20 mM potassium phosphate (pH 7.1)1.0 mM dithiothreitol50% (v/v) glycerol
Assay Conditions
50 mM Tris-HCl (pH 8.6 at 22°C)100 mM KCl10 mM MgCl21.0 mg/ml bovine serum albumin100 mM KCl1.0 mM dithiothreitol0.2 mM of each dCTP, dGTP, dTTP, and [α-32P]dATP15 μg activated DNAIncubation at 60°C for 30 minutesReaction volume of 50 μl
Quality Control
All preparations are assayed for contaminating endonuclease, exonuclease, and single- and double-stranded DNase activities
Storage Conditions
Store at -20ºCProduct shipped on dry ice
Downloads
Certificate of Analysis PDF-Current LotCertificate of Analysis PDF-Current LotCertificate of Analysis PDF-Current LotMSDS PDF
References
1. Stenesh, J. and Roe, B.A. (1972) Biochim. Biophys. Acta. 272, 156-1662. Hugh, G. and Griffin, M. (1994) PCR Technology, p.p. 228-2293. McClary, J. et al. (1991) J. DNA Sequencing and Mapping, p.p. 173-180
Certificate of Analysis PDF-Current LotCertificate of Analysis PDF-Current Lot