Bst DNA polymerase, Large Fragment
Cat. No.: SB-G3403-1600U
Size: 1600U
Description
his product is Bst DNA polymerase, Large Fragment, derived from Bacillus stearothermophilus , truncated and recombined by Escherichia coli, it has 5′→3′DNA polymerase activity and strong strand displacement activity, and lacks 5′→3′external nucleic acid Dicer activity. Bst DNA polymerase, Large Fragment is mainly used in isothermal amplification reactions, such as LAMP (Loop-mediated isothermal amplification), RCA (Rolling-circle amplification), etc.
- Source : The large fragment derived from Bacillus stearothermophilus DNA polymerase I was recombinantly expressed in Escherichia coli.
- Definition of enzyme activity : at 65°C, the amount of enzyme required to incorporate 10 nmol dNTP into the acid-insoluble precipitate within 30 min is defined as one enzyme activity unit.
- Purity and concentration : SDS-PAGE detection purity ≥ 95%; endogenous nucleic acid residue < 1 pg/μL (qPCR detection); 8 U/μL.
- Inactivation or inhibition : Heat at 80°C for 20 min.
- Enzyme storage buffer : 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X-100, 50% Glycerol, pH 7.1.
- 10 x Reaction Buffer : 200 mM Tris-HCl, 100 mM (NH 4 ) 2 SO 4 , 100 mM KCl, 20 mM MgSO 4 , 1% Triton X-100, pH 8.8.
Storage and Handling Conditions
Transport in wet ice packs; store at -20°C, valid for 12 months.
Composition
| Component Number |
Component |
G3403 |
| G3403-1 |
Bst DNA polymerase, Large Fragment |
200 μL |
| G3403-2 |
10 × Bst Reaction Buffers |
1mL |
| G3403-3 |
MgSO4 (100 mM) |
500 μL |
| Product Manual |
1 copy |
Steps
1. Isothermal amplification (LAMP):
| Component |
Volume |
| MgSO4 (100 mM) |
1.5 μL |
| dNTP Mix (10 mM) |
3.5 μL |
| FIP/BIP Primers (100 μM) |
0.4/0.4 μL |
| F3/B3 Primers (10 μM) |
0.5/0.5 μL |
| LoopF/LoopB Primers (10 μM) |
2/2 μL |
| Bst DNA polymerase, Large Fragment |
1 μL |
| 10× Bst Reaction Buffer |
2.5 μL |
| Template DNA |
1 μL |
| Nuclease Free Water |
To 25 μL |
Configure the reaction system according to the superscript (except Template DNA), mix gently and centrifuge briefly to precipitate the liquid.
2. Add the corresponding volume of template DNA to make the total volume 25 μL, gently blow and mix with a pipette, and centrifuge briefly.
3. Incubate the reaction system at a constant temperature of 60°C-65°C for 1 h.
4. According to different experimental requirements, Hydroxy naphthol blue or phenol red can be added to the LAMP reaction system for visual indication, and gel electrophoresis can also be used for detection and analysis.
Precautions
- Bst DNA polymerase, Large Fragment cannot be used for thermal cycle sequencing and PCR.
- Enzyme products should be placed in an ice box or ice bath when used, and should be stored at -20°C immediately after use. It is recommended to store in separate packages.
- For your safety and health, please wear a lab coat and disposable gloves for operation.
