Bst DNA Polymerase, Large Fragment

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 dithiothreitol
50% (v/v) glycerol

Assay Conditions

50 mM Tris-HCl (pH 8.6 at 22°C)
100 mM KCl
10 mM MgCl₂
1.0 mg/ml bovine serum albumin
100 mM KCl
1.0 mM dithiothreitol
0.2 mM of each dCTP, dGTP, dTTP, and [α-³²P]dATP
15 μg activated DNA
Incubation at 60°C for 30 minutes
Reaction 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 -20ºC
Product shipped on dry ice