Tanım
Hieff™ Super Bst DNA Polymerase is derived from Thermophilic Geobacillus sp. DNA Polymerase I and has been genetically engineered to eliminate its 5’→3’ exonuclease activity. This product exhibits strong 5’→3’ DNA polymerase activity, strand-displacement activity, and tolerance to dUTP, making it particularly suitable for contamination-controlled isothermal amplification reactions such as LAMP and CPA. Compared to Bst Plus DNA Polymerase, Hieff™ Super Bst DNA Polymerase demonstrates enhanced thermostability, which improves amplification performance. The enzyme is well-suited for low-template-concentration assays and high-temperature LAMP reactions (e.g., at 70°C), effectively enhancing detection specificity and shortening time-to-result. Its thermostable nature also enables the development of hot-start Bst formulations for further improvement in assay specificity.
Specifications
|
Cat NO. |
14411ES03 / 14411ES05 / 14411ES08 |
|
Size |
6,000 U / 24,000 U / 120,000 U |
Components
|
Components No. |
Name |
14411ES03 (6000 U) |
14411ES05 (24000 U) |
14411ES08 (120000 U) |
|
14411-A |
Hieff™ Super Bst DNA Polymerase(120 U/μL) |
50 μL |
200 μL |
1 mL |
|
14411-B |
10× Hieff™ Super Bst DNA Polymerase Buffer |
1 mL |
1 mL |
3 × 1 mL |
|
14411-C |
100 mM MgSO4 |
1 mL |
1 mL |
3 × 1 mL |
Applications
This product is suitable for various isothermal amplification methods, including LAMP, CPA, and RCA. It supports LAMP reactions at 70°C or, when combined with a thermostable reverse transcriptase, enables RT-LAMP at 70°C.
Unit Definition
One unit (U) is defined as the amount of enzyme that incorporates 10 nmol of dNTPs into acid-insoluble material 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
Store at –25 to –15°C for 2 years. Avoid repeated freeze-thaw cycles.
Notes
1. Keep the enzyme on ice or in an ice bath during use, and return it immediately to –20°C after use.
2. This product is for research use only.
3. For your safety and health, wear a lab coat and disposable gloves during handling.
LAMP Experimental Example
1. Reaction Setup
|
Component |
Volume (μL) |
Final Concentration |
|
10× Hieff™ Super Bst DNA Polymerase Buffer |
2.5 |
1× |
|
100 mM MgSO₄ |
0.75 |
3 mM + 2 mM in buffer = 5 mM |
|
dNTP Mix (25 mM each) |
1.4 |
1.4 mM each |
|
dUTP (25 mM) (optional) |
1.4 |
1.4 mM |
|
UDGase (1 U/μL) (optional) |
1 |
0.04 U |
|
Template DNA |
10 ng–1 μg |
— |
|
10× Primers |
2.5 |
— |
|
Hieff™ Super Bst DNA Polymerase (120 U/μL) |
0.34–1* |
— |
|
ddH₂O |
to 25 |
— |
[Note]: The amount of Hieff™ Super Bst DNA Polymerase may be optimized according to specific experimental requirements.
10× Buffer composition: 200 mM Tris-HCl, 500 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, 1% Tween-20, pH 8.8 @ 25°C.
Mg²⁺ concentration can be adjusted between 4–10 mM depending on the assay.
10× Primers: 16 µM FIP/BIP, 2 µM F3/B3, 4 µM Loop F/B each.
Our dNTP (Cat#10124), dUTP (Cat#10128), and UDGase (Cat#10303) are compatible with this product.
2. Reaction Conditions
|
Temperature |
Time |
Purpose |
|
25–37°C |
5–10 min |
Degradation of uracil-containing templates (optional) |
|
65–70°C |
30–60 min |
Amplification |
|
85°C |
5 min |
Enzyme inactivation |
Documents:
Safety Data Sheet
Manuals
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