Bst 2.0 DNA Polymerase(Glycerol free)
Bst DNA polymerase V2 is derived from Bacillus stearothermophilus DNA Polymerase I, which has 5′→3′ DNA polymerase activity and strong chain replacement activity, but no 5′→3′ exonuclease activity. Bst DNA Polymerase V2 is ideally suitable for strand-displacement, isothermal amplification LAMP (Loop mediated isothermal amplification) and rapid sequencing.
Components
|
Component |
HC5005A-01 |
HC5005A-02 |
HC5005A-03 |
|
BstDNApolymerase V2(Glycerol-free)(8U/μL) |
0.2 mL |
1 mL |
10 mL |
|
10×HC Bst V2 Buffer |
1.5 mL |
2×1.5 mL |
3×10 mL |
|
MgSO4 (100mM) |
1.5 mL |
2×1.5 mL |
2×10 mL |
Applications
1.LAMP isothermal amplification
2.DNA strand single displacement reaction
3.High GC gene sequencing
4.DNA sequencing of nanogram level.
Storage Condition
Transportation under 0°C and be stored at -25°C~-15°C.
Unit Definition
One unit is defined as the amount of enzyme that incorporate 25 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
Quality Control
1.Protein Purity Assay (SDS-PAGE): The purity of Bst DNA polymerase V2 is ≥99% determined by SDS-PAGE analysis using Coomassie Blue detection.
2.Exonuclease Activity: Incubation of a 50 μL reaction containing a minimum of 8 U of Bst DNA polymerase V2 with 1 μg λ -Hind Ⅲ digest DNA for 16 hours at 37 ℃ results in no detectable degradation as determined.
3.Nickase Activity: Incubation of a 50 μL reaction containing a minimum of 8 U of Bst DNA polymerase V2 with 1 μg pBR322 DNA for 16 hours at 37°C results in no detectable degradation as determined.
4.RNase Activity: Incubation of a 50 μL reaction containing a minimum of 8 U of Bst DNA polymerase V2 with 1.6 μg MS2 RNA for 16 hours at 37°C results in no detectable degradation as determined.
5.E. coli DNA: 120 U of Bst DNA polymerase V2 is screened for the presence of E. coli genomic DNA using TaqMan qPCR with primers specific for the E. coli 16S rRNA locus. The E. coli genomic DNA contamination is ≤1 Copy.
LAMP Reaction
|
Components |
25 μL |
|
10×HC Bst V2 Buffer |
2.5 μL |
|
MgSO4 (100mM) |
1.5 μL |
|
dNTPs (10mM each) |
3.5 μL |
|
SYTO™ 16 Green (25×) a |
1.0 μL |
|
Primer mix b |
6 μL |
|
Bst DNA Polymerase V2 (Glycerol-free) (8 U/uL) |
1 μL |
|
Template |
× μL |
|
ddH₂O |
Up to 25 μL |
Notes:
1) a. SYTOTM 16 Green (25×): According to experimental needs, other dyes can be used as substitutes;
2) b. Primer mix: obtained by mixing 20 µ M FIP, 20 µ M BIP, 2.5 µ M F3, 2.5 µ M B3, 5 µ M LF, 5 µ M LB and other volumes.
Reaction and Condition
1 × HC Bst V2 Buffer, the incubation temperature is between 60°C and 65°C.
Heat Inactivation
80 °C，20mins
