SKU : G3403-1600U
หมวดหมู่ : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
แบรนด์ : Servicebio
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Product Information
Product Name | Cat. No. | Spec.. |
Bst DNA polymerase, Large Fragment | G3403-1600U | 1600 U |
Product Description/Introduction
Bst DNA Polymerase, Large Fragment is the large fragment of Bacillus stearothermophilus (Bst) DNA Polymerase I with 5'3' DNA polymerase activity, but not 5'3' exonuclease activities. Bst DNA Polymerase, Large Fragment has strong strand displacement ability and can be applied to isothermal nucleic acid amplification reactions, such as Loop-mediated isothermal amplification (LAMP) and Rolling-circle amplification (RCA), etc.
Applications: Loop-mediated isothermal amplification (LAMP), helicase isothermal gene amplification (HDA) and other DNA isothermal amplification; multiple displacement amplification (MDA); whole genome amplification (WGA); high GC content DNA sequencing; rapid sequencing of nanogram-level DNA templates; library preparation for sequencing, etc.
Source: The recombinant large fragment of Bacillus stearothermophilus DNA Polymerase I expressed in E. coli.
Purity and concentration: Purity 95% by SDS-PAGE; endonuclease ＜1 pg/μLby qPCR; 8 U/uL.
Definition of activity: One activity unit is defined as the amount of enzyme that can incorporate 10nmol of dNTPs into acid insoluble material in 30 minutes at 65.
Enzyme storage buffer: 10mM Tris-HCl (pH 7.5), 50mM KCl, 0.1mM EDTA, 1mM DTT, 0.1% Triton X-100, 50% (v/v) glycerol.
10X Bst Reaction Buffer: 200mM Tris-HCl, 100mM KCl, 100mM (NH4)2SO4 , 20mM MgSO4 , 1% Triton X-100 , pH 8.8 at 25°C.
Inactivation or inhibition: Heat at 80 for 20min.
Storage and Shipping Conditions
Ship with wet ice; store at -20°C, valid for 12 months.
Product Content
Component Number | Component | G3403-1600U |
G3403-1 | Bst DNA polymerase, Large Fragment | 200 μL |
G3403-2 | 10×Bst Reaction Buffer | 1 mL |
G3403-3 | MgSO4 (100 mM) | 500 μL |
Manual | One copy | |
Assay Protocol / Procedures
Taking LAMP isothermal amplification as an example.
1. Set up the following reaction.
Component | Volume |
10× Bst Reaction Buffer | 2.5 μl |
MgSO4 (100 mM) | 1.5 μl |
dNTP Mix (10 mM each) | 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 |
Template DNA | 1 μL |
Nuclease-free Water | To 25 μL |
2. Incubate at 60-65 for 1 hour.
3. According to different experimental requirements, the LAMP reaction system can be added with hydrogen naphthol blue or phenol red for visual indication, or gel electrophoresis can be used for detection and analysis.
Note
1. Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing and PCR.
2. When using enzyme products, they should be placed on ice. After use, they should be immediately stored at -20 , and it is recommended to store them separately.
3. For your safety and health, please wear a lab coat and disposable gloves during the operation.
For Research Use Only!
