Bst DNA polymerase, large fragment
A large fragment of DNA polymerase Bacillus stearothermophilus. The enzyme is highly processive and catalyzes DNA synthesis in the 5'-3' direction.
Catalog #
Quantity, u.a.
Qty
Bst DNA polymerase, large fragment, is a single polypeptide with a molecular weight of about 67 kDa. The polymerase is highly processive. Bst DNA polymerase catalyzes DNA synthesis in the 5'-3' direction. The enzyme lacks 5'-3' and 3'-5' exonuclease activity, but has 5'-3' displacing activity. Bst DNA polymerase was purified from an E. coli strain containing a plasmid with a cloned full-length gene of the large fragment of Bacillus stearothermophilus DNA polymerase I. The enzyme contains bacterial DNA.
One unit of activity is the amount of enzyme required to incorporate 10 nmol of dNTP into the acid-insoluble DNA fraction in 30 min at 65°C, pH 8.8.
Quality control. Each batch of enzyme is tested for the absence of endonuclease and non-specific exonuclease activity, and enzyme sensitivity.
One unit of activity is the amount of enzyme required to incorporate 10 nmol of dNTP into the acid-insoluble DNA fraction in 30 min at 65°C, pH 8.8.
Quality control. Each batch of enzyme is tested for the absence of endonuclease and non-specific exonuclease activity, and enzyme sensitivity.
Application
PCR isothermal
Advantages
Bst DNA polymerase is more resistant to inhibitors, has higher specific activity and processivity than Taq DNA polymerase.
Properties
Typical LAMP protocol:
| Component | Final concentration |
| 10X Buffer for Bst | 1X (does not contain MgSO4) |
| 8 mM | |
| dNTP Mix (10 mM) | 1.4 mM each |
| FIP/BIP Primers (10X) | 1.6 µM |
| F3/B3 Primers (10X) | 0.2 µM |
| < p> LoopF/B Primers (10X) | 0.8 µM |
| Bst DNA polymerase (16,000 U/ml) | 80-320 U/ml |
| DNA sample | > 10 copies per reaction |
| Nuclease-free water | up to 25 µl |
| Total reaction volume | 25 µl |
Storage conditions
-20°C in 10 mM Tris-HCl (pH 8.0 at 25°C), 10 mM KCl, 1% BSA, 0.02% Tween 20, 50% glycerol.
