Bst DNA Polymerase (Large Fragment)
Bst DNA Polymerase (Large fragment) is an enzyme of Bacillus stearothermophilus DNA polymerase which can catalyze 5´ → 3´ polymerase activity but lacks 5´ →3´ exonuclease activity. Bst DNA Polymerase offers strand displacement capabilities, making it ideal for isothermal amplification.
Package & Component :
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE (purified by Ni-NTA chromatography).
Unit Definition:
One unit is defined as the amount of the enzyme incorporates 10 nmol of dNTP into acid-insoluble product in 30 minutes at 65°C.
Reaction Condition:
1X Bst DNA Polymerase reaction buffer, supplemented with dNTP mix, primer and DNA template. Incubate at 65°C.
10X Bst DNA Polymerase Reaction Buffer: 200 mM Tris-HCl (pH 8.8), 100 mM (NH4)2SO4, 100 mM KCl, 20 mM MgSO4, and 1% Triton® X-100.
Storage Buffer:
Bst DNA Polymerase (Large fragment) is supplied in 10 mM Tris-HCl (pH7.5), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton® X-100 and 50% Glycerol.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Notes:
It is not recommended to perform reaction above 70 °C. Bst DNA Polymerase cannot be used for thermal cycle sequencing
Shipping Conditions:
Blue ice
| Cat. | Name | Amount |
| C15019-1600U | Bst DNA Polymerase (Large Fragment) (8 U/μL) | 1,600 U |
| 10X Bst DNA Polymerase Reaction Buffer | 1 mL | |
| 100 mM MgSO4 | 0.4 mL | |
| C15019-8000U | Bst DNA Polymerase (Large Fragment) (8 U/μL) | 8,000 U |
| 10X Bst DNA Polymerase Reaction Buffer | 1 mL | |
| 100 mM MgSO4 | 0.4 mL |
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE (purified by Ni-NTA chromatography).
Unit Definition:
One unit is defined as the amount of the enzyme incorporates 10 nmol of dNTP into acid-insoluble product in 30 minutes at 65°C.
Reaction Condition:
1X Bst DNA Polymerase reaction buffer, supplemented with dNTP mix, primer and DNA template. Incubate at 65°C.
10X Bst DNA Polymerase Reaction Buffer: 200 mM Tris-HCl (pH 8.8), 100 mM (NH4)2SO4, 100 mM KCl, 20 mM MgSO4, and 1% Triton® X-100.
Storage Buffer:
Bst DNA Polymerase (Large fragment) is supplied in 10 mM Tris-HCl (pH7.5), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton® X-100 and 50% Glycerol.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Notes:
It is not recommended to perform reaction above 70 °C. Bst DNA Polymerase cannot be used for thermal cycle sequencing
Shipping Conditions:
Blue ice
LAMP reaction recipe:
1. Place all required reagents on ice and add each of them following the order suggested below.
2. Gently mix the reaction thoroughly to achieve uniform distribution.
3. Incubate at 65°C for 30-60 minutes.
4. MgSO4 (2-10 mM), Bst DNA Polymerase (40-320 U/mL) and temperature (50-65 °C) can be adjusted for optimal results.
5. Reaction preparations may be scaled up or down proportionately.
1. Place all required reagents on ice and add each of them following the order suggested below.
| Component | Amount | Final concentration |
| 10X Bst DNA Polymerase Reaction Buffer | 2.5 μL | 1X |
| 100 mM MgSO4 | 1.5 μL | 6 mM final concentration, total 8 mM |
| 10 mM dNTP mix | 3.5 μL | 1.4 mM each |
| 10X FIP/BIP primers | 1 μL | 1.6 μM |
| 10X F3/B3 primers | 1 μL | 0.2 μM |
| 10X LoopF/B primers | 1 μL | 0.8 μM |
| DNA template | X μL | 10 copies or more |
| Nuclease-Free H2O | Y μL | - |
| Bst DNA Polymerase (Large Fraction) (8 U/μL) |
1 μL | 8 U/rxn |
| Total reaction volume | 25 μL | - |
3. Incubate at 65°C for 30-60 minutes.
4. MgSO4 (2-10 mM), Bst DNA Polymerase (40-320 U/mL) and temperature (50-65 °C) can be adjusted for optimal results.
5. Reaction preparations may be scaled up or down proportionately.
