Bst 2.0 DNA Polymerase

Description

Bst DNA polymerase is a homology of Bacillus Stearothermophilus DNA polymerase I, large fragment (Bst DNA polymerase, large fragment) and was designed by computer simulation. BstL DNA polymerase has 5´→3´polymerase activity and strong chain substitution activity but does not have 5´→3´ exonuclease activity. Compared with wild-type Bst DNA polymerase, the enzyme can effectively increase the amplification speed, yield, salt tolerance, and thermal stability, etc

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

Application

1. Isothermal amplification.
2. DNA sequencing with high GC content
3. Rapid Sequencing from nanogram amounts of DNA template.

Storage Conditions

Transport in ice packs, store at -20℃. Avoid repeated freezing and thawing. The product is valid for 1 years.

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

Exonuclease residue detection: The 50 μL reaction system added 16 U enzyme, 1 μg of PUC19 DNA vector, and incubated at 37℃ for 4 hours without degradation signal.

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

RNase residue detection RNase: 16 U enzyme and 40 ng RNA (3500 nt) 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 16 U of this enzyme was detected by TaqMan qPCR for E. coli gDNA, and the E. coli genome residue was less than 1 copy.

Experiment Process 1.Take out 10×BstL Buffer, thaw on ice, 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.

3. Mix by pipetting, and centrifuge briefly to collect.

4. Constant temperature incubation at 60°C- 65°C for 1 hour.

Precautions

1) BstL Buffer can be dissolved at room temperature. After dissolution, it should be stored in an icebox or ice bath. After using, it should be stored at -20℃ immediately.

2) For your safety and health, please wear a lab coat and disposable gloves.