Bst DNA polymerase (large fragment)​

Bst DNA Polymerase Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the 5´→3´polymerase activity, but lacks 5´→3´exonuclease activity. This product uses E. coli expression and is suitable for isothermal amplification.

Store Buffer

10 mM Tris-HCl (pH 7.1)，50 mM KCl， 0.1 mM EDTA，1 mM DTT，0.1% Triton X-100，50% Glycerin.

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

Quality Control

Endonuclease residue detection: The 50 μL reaction system added 500 U enzyme, 1 μg of PUC19 DNA, and was incubated at 37℃ for 4h without degradation signal.

Exonuclease residue detection: After incubation at 37℃ for 16 h, the electrophoretic bands of DNA did not change after adding 500 U enzyme and 1 μg Lambda-HindIII DNA into the reaction system.

RNase residue detection RNase: 8 U enzyme and 40 ng RNA were incubated at 37℃ for 16 h, and the electrophoresis band of RNA did not change.

E. coli DNA residue detection: The remaining nucleic acid in 120 U of this enzyme was detected by TaqMan qPCR for E. coli gDNA, and the E. coli genome residue was less than 10 copies.

Experiment Process

1) Take out 10×Bst Buffer, thaw on ice, and vortex for 10 seconds before use. Mix well and collect by centrifugation briefly.

2) In addition to the template DNA, the remaining components are configured in the order shown in the table below.

 

1) Isothermal amplification.

2) DNA sequencing with high GC content.

3) Rapid Sequencing from nanogram amounts of DNA template.