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Bst DNA Polymerase with Strand Displacement Activity


For Isothermal Amplification, Whole Genome Amplification, MDA, and DNA Sequencing
• High specific activity and high purity
• Enhanced strand displacement activity
• Suitable for isothermal amplification, whole genome amplification (WGA), multiple displacement amplification (MDA), and next generation sequencing
• Available in two concentrations: 8 U/ul and 50 U/ul

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Lucigen´s Bst DNA Polymerase (Exonuclease Minus) possesses more strand displacement activity than those from other suppliers.
M13 single stranded DNA was incubated with or without 8 units of Bst DNA Polymerase (±Bst) in the reaction buffer supplied by the manufacturer in the presence or absence of replication primer ± primer) for 30 minutes at 65°C.
MW: 1 kb ladder.


Advantages
The properties of this enzyme can be applied to nucleic acid amplification methods, including isothermal amplification, whole genome amplification (WGA), multiple displacement amplification (MDA), and is useful in next generation sequencing.
Expressed in E. coli, this enzyme has optimal activity at 65°C, and is suitable for sequencing DNA with high GC content and secondary structures. It is available in two concentrations: 8 U/ul and 50 U/ul.

Description
Bst DNA Polymerase (Exonuclease Minus), is a 67 kDa Bacillus stearothermophilus DNA Polymerase protein (large fragment) which has 5’-3’ polymerase activity and strand displacement activity but lacks 3’-5’ exonuclease activity. It also has reverse transcription activity.

Bst DNA Polymerase, Exonuclease Minus, is supplied with 10X DNA Polymerase Buffer B, composed of 200 mM Tris-HCl pH 8.8, 100 mM (NH4)2SO4, 100 mM KCl, 20 mM MgSO4, and 1.0 % Triton X-100.

Purity: >99% pure by SDS-PAGE. No detectable DNA contamination. 10 µl of enzyme at 8 U/µl of the sample was tested for E. coli genomic DNA contamination by PCR amplifying with the E. coli 16S ribosomal primers.

Activity Determination: One unit catalyzes the incorporation of 10 nmol of dNTP into acid-insoluble material in 30 minutes at 65°C in 20 mM Tris-HCl pH 8.8, 10 mM (NH4)2SO4, 10 mM KCl, 2 mM MgSO4, 0.1 % Triton X-100, 30 nM M13mp18 ssDNA, 70 nM M13 sequencing primer(-47) 24 mer 200 µM dGTP, dATP, dTTP, dCTP (a mix of unlabeled and and 33P]dCTP), and 0.1 mg/ml BSA.

Absence of Endonuclease or Nicking Activity: Incubation of 8 U and 50 U of Bst DNA Polymerase, Exonuclease Minus, with 1 µg of supercoiled pBR322 DNA for 16 hours at 37º and 65ºC resulted in no detectable conversion to relaxed or linear forms by agarose gel electrophoresis.

Absence of Exonuclease Activity: Incubation of 8 U and 50 U of Bst DNA Polymerase, Exonuclease Minus, with 1 µg of HindIII-cut lambda DNA for 16 hours at 37º and 65ºC resulted in no smearing of bands on agarose gels. Single stranded and double stranded exonuclease activities were tested by incubating 10 µl of enzyme at 8 U/µl with radiolabeled DNA substrate for one hour at 37º and 65ºC, resulting in less than 0.1% release of TCA-soluble counts.

Applications:
• Strand displacement amplification
• DNA sequencing through high GC regions (1,2)
• Rapid sequencing from nanogram amounts of DNA template (3)

References
(1) Griffin, H. and Griffin, A. (1994) PCR Technology, 228-229.
(2) McClary, J. et al. (1991) J. DNA Sequencing and Mapping, 1, 173-180.
(3) Mead, D.A. et al. (1991) Biotechniques, 11, 76-87.
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Description Cat# Size Price    
Bst DNA Polymerase (Exo minus) (50 U / ul) 30028-1-LU 10000 units 404 € DETAILS   Add to Cart 
Bst DNA Polymerase (Exo minus) (8 U / ul) 30027-1-LU 2000 units 160 € DETAILS   Add to Cart 
Bst DNA Polymerase (Exo minus) (8 U / ul) 30027-2-LU 10000 units (5 x 2000 units) 404 € DETAILS   Add to Cart 
Bst DNA Polymerase, Large Fragment M1213-200-BV 1600 U (200 ul) 265 € DETAILS   Add to Cart 

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