Features
Thermophilic DNA polymerase with strong strand displacement activity.
Concentration
8u/µl
Applications
- Isothermal DNA amplification by the method of:
– loop-mediated isothermal amplification (LAMP) (1, 2),
– whole genome amplification (WGA) (3),
– ramification amplification (RAM) (4).
- Random-primed DNA labeling.
- Labeling by fill-in 5’-overhangs of dsDNA.
Description
Bsm DNA Polymerase, Large Fragment, is a portion of DNA polymerase of Bacillus smithii, which catalyzes 5'=>3' synthesis of DNA and lacks 5'=>3' and 3'=>5' exonuclease activities. Bsm DNA Polymerase, Large Fragment, has a strong strand displacement activity and is active in a wide range of temperatures from 30°C to 63°C, with an optimum of activity at 60°C. Bsm DNA Polymerase, Large Fragment, is an enzyme with high functional similarity to Bst DNA Polymerase, Large Fragment, and can replace it in most applications.
Not suitable for use in PCR.
Source
E.coli cells with a cloned polA gene from Bacillus smithii.
Molecular Weight
67 kDa monomer.
Definition of Activity Unit
One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 60°C.
Enzyme activity is assayed in the following mixture: 10 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 100 µg/ml BSA, 0.75 mM activated salmon milt DNA, 0.2 mM of each dNTP, 0.4 MBq/ml [H3]-dTTP.
Storage Buffer
The enzyme is supplied in:
10 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.15% (v/v) Triton X-100, 50% (v/v) glycerol.
10X Bsm Buffer
200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM KCl, 100 mM (NH4)2SO4, 20 mM MgSO4, 1% (v/v) Tween 20