BspQI restriction endonuclease
BspQI can be recombinantly expressed in E. coli that can recognize specific sites and are produced under
BspQI, an IIs restriction endonuclease restriction endonuclease, is derived from a recombinant E. coli strain that carries the cloned and modified BspQI gen from Bacillus sphaericus. It can recognize specific sites, and recognition sequence and cleavage sites are as follows:
5' · · · ·GCTCTTC(N) · · · · · · · · · · ·3'
3' · · · · CGAGAAG(NNNN) · · · · 5'
Product Features
1. High activity, Fast digestion;
2. Low star activity, ensuring accurate cutting like “scalpel”;
3. Without BSA and animal-origin free;
Methylation Sensitivity
Dam methylation: Not Sensitive;
Dcm methylation: Not Sensitive;
CpG Methylation: Not Sensitive;
Storage conditions
The product should be shipped ≤ 0℃; Store at -25~- 15℃ condition.
Storage buffer
20mM Tris-HCl, 0.1mM EDTA, 500 mM KCl, 1.0 mM dithiothreitol, 500 µg/ml Recombinant Albumin, 0. 1% Trition X- 100 and 50% glycerol (pH 7.0 @ 25°C).
Unit Definition
One unit is defined as the amount of enzyme required to digest 1µg of Internal control DNA in 1 hour at 50°C in a total reaction volume of 50 µL.
Quality Control
Protein Purity Assay (SDS-PAGE): The purity of BspQI was ≥95% determined by SDS-PAGE analysis.
RNase: 10U of BspQI with 1.6μg MS2 RNA for 4 hours at 50℃ yields no degradation as determined by agarose gel electrophoresis.
Non-Specific DNase Activity: 10U of BspQI with 1μg λ DNA at 50℃ for 16 hours, compared with 50℃ for 1hour, yields no excess DNA as determined by agarose gel electrophoresis.
Ligation and Recutting: After digestion of 1 μg λDNA with 10U BspQI, DNA fragments can be ligated with T4 DNA ligase at 16ºC. And these ligated fragments can be recut with BspQI.
E. coli DNA: E. coli 16s rDNA-specific TaqMan qPCR detection showed that E.coli genome residue ≤ 0.1pg/ul.
Host protein residue: ≤ 50 ppm
Bacterial Endotoxin: LAL-test, according to Chinese Pharmacopoeia IV 2020 edition, gel limit test method, general rule (1143). Bacterial endotoxin content should be ≤10 EU/mg.
Reaction system and conditions
|
Component |
Volume |
|
BspQ I(10 U/μL) |
1 μL |
|
DNA |
1 μg |
|
10 x BspQ I Buffer |
5 μL |
|
dd H2O |
Up to 50 μL |
Reaction conditions：50℃, 1~ 16 h.
Heat inactivation：80°C for 20 min.
The recommended reaction system and conditions can provide relatively good enzyme digestion effect,which is for reference only, please refer to the experimental results for details.
Product application
Restriction endonuclease digestion, Rapid cloning.
Notes
1. The volume of enzyme ≤ 1/10 of the reaction volume.
2. Star activity may occur when glycerol concentration is more than 5%.
3. Cleavage activity may occur when Substrate below the recommended ratio.
