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Bsm DNA Polymerase, Large Fragment

FastDigest buffer for 100% activity G buffer for 100% activity LO certified O buffer for 100% activity Recombinant enzyme R buffer for 100% activity Tango buffer for 100% activity Thermal inactivation at 80°C in 10 min

DNA polymerase with a high functional similarity to Bst DNA Polymerase and with a strong strand displacement activity.
  
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Thermo ScientificBsm 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°. It 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.

Highlights

  • Thermophilic DNA polymerase with strong strand displacement activity

Applications

  • Isothermal DNA amplification by the method of:
    • Loop-mediated isothermal amplification (LAMP) (see Reference1, 2)
    • Whole genome amplification (WGA) (see​ Reference3)
    • Ramification amplification (RAM) (see​ Reference4)
     
  • Random-primed DNA labeling
  • Labeling by fill-in 5'-overhangs of dsDNA

Note

Use of this enzyme in certain applications may be covered by patents and may require a license.

  
Storage Condition-20 C
HazardousNo
10X Bsm Buffer200mM Tris-HCl (pH8.8 at 25°C), 100mM KCl, 100mM (NH4)2SO4, 20mM MgSO4, 1%(v/v) Tween 20.
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.
 
InactivationInactivated by heating at 80°C for 10min.
Molecular Weight67kDa monomer
Quality ControlThe absence of endodeoxyribonucleases, exodeoxyribonucleases, ribonucleases, and E.coli genomic DNA is confirmed by appropriate quality test.
SourceE.coli cells with a cloned polA gene from Bacillus smithii.
Storage BufferThe enzyme is supplied in 10mM Tris-HCl (pH7.5), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.15%(v/v) Triton X-100, 50%(v/v) glycerol.

References

  1. N. Tsugunori et al., Loop-mediated isothermal amplification of DNA.Nucleic Acids Res.28(12), e63 (2000).
  2. I. Masaki et al., Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemaglutinin gene-specific loop-mediated isothermal amplification method.Journal of Virological Methods.141, 173-180 (2007).
  3. F. B. Dean et al., Comprehensive human genome amplification using multiple displacement amplification.Proc. NatI. Acad. Sci. USA.99, 5261-5266 (2002).
  4. Y. Jizu et al., Molecular Zipper: a fluorescent probe for real-time isothermal DNA amplification.Nucleic Acids Res.34(11), e81 (2006).