説明
Bst DNA Polymerase is derived from Bacillus stearothermophilus DNA Polymerase I. It possesses 5'→3' DNA polymerase activity and strong strand displacement activity but lacks 5'→3' exonuclease activity. It is suitable for DNA strand displacement reactions, isothermal amplification techniques such as LAMP (Loop-mediated Isothermal Amplification), and rapid sequencing.
WS Bst DNA Polymerase is a hot-start version of Bst DNA Polymerase developed using reversible modification technology. It effectively inhibits DNA polymerase activity at room temperature, allowing for reaction setup and operation at ambient conditions without non-specific amplification, thereby improving reaction efficiency. Furthermore, the enzyme releases its activity at elevated temperatures, eliminating the need for a separate activation step.
Specifications
|
Cat.No. |
14412ES03 / 14412ES05 / 14412ES08 |
|
Size |
8,000 U / 40,000 U / 400,000 U |
Components
|
Components No. |
Name |
14412ES03 (8,000 U) |
14412ES05 (40,000 U) |
14412ES08 (400,000 U) |
|
14412-A |
HieffTM WS Bst DNA Polymerase(40 U/μL) |
200 μL |
1 mL |
10 mL |
|
14412-B |
10× HieffTM WS Bst DNA Polymerase Buffer |
1 mL |
3 × 1 mL |
3 × 10 mL |
|
14412-C |
100 mM MgSO4 |
1 mL |
1 mL |
10× 1 mL |
Product Applications
This product is suitable for various isothermal amplification reactions, including LAMP, CPA, and RCA.
Unit Definition
1 Unit (U) is defined as the amount of enzyme required to incorporate 10 nmol of dNTP into acid-insoluble precipitates in 30 minutes at 65°C.
Enzyme Storage Buffer Composition
10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% Glycerol, pH 7.5 @ 25°C.
Storage
This product should be stored at -25~-15℃ for 2 years.Avoid repeated freeze-thaw cycles.
Notes
1. When in use, the enzyme should be kept on ice or in an ice bath. Immediately after use, store it at -20°C.
2. This product is for research use only.
3. Please operate with lab coats and disposable gloves，for your safety.
LAMP Experimental Case Study
1. Reaction System Preparation
|
Component |
Volume (μL) |
Final Concentration |
|
10× HieffTM WS Bst DNA Polymerase Buffer |
1× |
|
|
1.4 mM each |
||
|
dUTP (25 mM) (Optional addition) |
1.4 |
1.4 mM |
|
UDGase (1 U/μL) (Optional addition) |
1 |
0.04 U |
|
Template DNA |
10 ng~1 μg |
- |
|
10×Primers |
2.5 |
- |
|
HieffTM WS Bst DNA Polymerase(40 U/μL) |
1* |
- |
|
ddH2O |
to 25 |
- |
【Notes】:
1)The amount of HieffTM WS Bst DNA Polymerase can be adjusted and optimized depending on the specific application.
2)The Mg²⁺ concentration can be adjusted between 8–10 mM depending on the experiment. The final Mg²⁺ concentration is 8 mM when using the 10× buffer.
3)10× Primers: 16 µM FIP/BIP, 2 µM F3/B3, and 4 µM each for Loop F and Loop B.
4)dNTP (Cat#10124), dUTP (Cat#10128), and UDGase (Cat#10303) from our company are compatible with this product.
2. Reaction Conditions
|
Temperature |
Time |
Function |
|
25~37°C |
5~10 min |
Degradation of U-containing templates (optional) |
|
60~65°C |
30~60 min |
Reaction |
|
85℃ |
5 min |
Inactivation |
Documents：
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