BstDNA Polymerase 1.0
Bst DNA Polymerase 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.
10 mM Tris-HCl (pH 7.1)，50 mM KCl，0.1 mM EDTA，1 mM DTT，0.1% Triton X-100，50% Glycerin.
- Isothermal amplification.
- DNA sequencing with high GC content.
- Rapid Sequencing from nanograms of DNA template.
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.
- Endonuclease residue detection: The 50 μL reaction system added 500 U enzyme, 1 μg of PUC19 DNA, and 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.
- Take out 10×Bst Buffer, thaw on ice, vortex for 10 seconds before use. Mix well and collect by centrifugation briefly.
- In addition to the template DNA, the remaining components are configured in the order shown in the table below.
|10 ×Bst Buffer||2.5 μL||1 ×(2 mM Mg2+ )|
|MgSO4 (100 mM)||1.5 μL||6 mM (total 8 mM)|
|dNTP Mix (10 mM)||3.5 μL||1.4 mM|
|FIP/BIP Primers (25×)||1 μL||1.6 μM|
|F3/B3 Primers (25×)||1 μL||0.2 μM|
|LoopF/LoopB Primers||1 μL||0.4 μM|
|Bst DNA Polymerase（8 U/μL）||1 μL||320 U/mL|
|Template DNA||x μL||>10 copies|
|Nuclease-free Water||up to 25 μL||—|
3. Mix by pipetting, and centrifuge briefly to collect. 4. Constant temperature incubation at 60°C- 65°C for 1 hour.
- The Bst Buffer can be dissolved at room temperature. After dissolution, it should be stored in an ice box or ice bath. After using, it should be stored at -20℃ immediately.
- For your safety and health, please wear lab coat and disposable gloves.