Beskrivelse
Bst Plus DNA Polymerase is derived from Bacillus stearothermophilus DNA Polymerase I and has been genetically engineered to eliminate its 5’→3’ exonuclease activity while retaining robust 5’→3’ DNA polymerase activity and strong strand-displacement capability. Compared with wild-type Bst DNA Polymerase, this enzyme demonstrates significant improvements in amplification speed, yield, salt tolerance, and thermostability. Additionally, it exhibits enhanced dUTP tolerance, making it highly suitable for isothermal amplification applications such as LAMP (Loop-Mediated Isothermal Amplification). This product is glycerol-free Bst Plus DNA Polymerase, ideal for lyophilized formulation development.
Specifications
|
Cat NO. |
14413ES60 / 14413ES97 / 14413ES98 |
|
Size |
12 KU / 120 KU / 1,200 KU |
Components
|
Name |
14413ES60 |
14413ES97 |
14413ES98 |
|
Bst Plus DNA Polymerase (60 U/μL，Glycerol-Free) |
200 μL |
2 mL |
20 mL |
Applications
Isothermal amplification reactions.
Unit Definition
One unit (1 U) is defined as the amount of enzyme required to incorporate 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 65°C.
Heat Inactivation
85°C for 5 minutes.
Storage
Shipped at ambient temperature protected from light. Store at –20°C protected from light. Stable for 1 year.
Notes
1. For your safety and health, please wear a lab coat and disposable gloves during handling.
Instructions: LAMP Assay Using pH Indicator Method
1. Reaction Setup
|
Component |
Volume per Reaction (μL) |
Final Concentration |
|
2.5× pH Sensitive Reaction Buffer |
10 |
1× |
|
1 M MgSO₄ |
0.2 |
— |
|
25 mM d(A/U/G/C)TPa |
1.4 |
— |
|
UDGase |
0.005 |
— |
|
Glycerol-Free Reverse Transcriptase |
0.075 |
— |
|
Bst Plus DNA Polymerase |
0.67 |
— |
|
Glycerol-Free RNase Inhibitor |
0.2 |
— |
|
10× Primersb |
2.5 |
1× |
|
Template |
10 ng – 1 μg |
— |
|
ddH₂O |
Up to 25 |
— |
[Note]: a. The recommended final concentration of the 25 mM d(A/U/G/C)TP mix is 1.2–1.4 mM.
b. 10× Primer Mix composition: 16 µM FIP/BIP, 2 µM F3/B3, and 4 µM each of Loop F/Loop B; primer ratios may be adjusted as needed.
2. Reaction Conditions
Incubate at 65°C for 30–60 minutes, followed by heat inactivation at 85°C for 5 minutes.
If using a UDG system, include a UDG digestion step (e.g., 25°C for 10 minutes) prior to amplification.
Documents:
Safety Data Sheet
Manuals
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Forespørgsel
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FAQ
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