BspQ I restriction endonuclease HYJ111
BspQ I restriction endonuclease is a recombinant protein encoded by the BspQ I gene of Bacillus sphaericus expressed recombinantly in Escherichia coli. It is a class IIS restriction endonuclease that recognizes specific sites. Its recognition sequence and cleavage site are as follows:
5'······GCTCTTC(N)··············3'
3'······CGAGAAG(NNNN)······5'
Components
|
Components |
0.5KU |
2.5KU |
10KU |
100KU |
1000KU |
|
BspQ I (10 U/μL) |
50 uL |
250 uL |
1 mL |
10 mL |
100 mL |
|
10×BspQ I Buffer |
1.25 mL |
1.25 mL |
4 × 1.25 mL |
5 × 10 mL |
5 × 100 mL |
Storage
Store at -25°C to -15°C, valid for 2 years.
Storage buffer
20 mM Tris-HCl, 0.1 mM 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).
Activity Definition
One unit of activity is defined as the amount of enzyme required to completely cleave 1 μg of internal control DNA in a 50 μL reaction at 50°C within 1 hour.
Quality control
1. Protein purity (SDS-PAGE): BspQ I purity was ≥95% as determined by SDS-PAGE analysis.
2. RNase activity: 10 U of this product reacted with 1.6 μg of MS2 RNA at 50°C for 4 hours, resulting in no change in the RNA electrophoretic bands.
3. Nonspecific DNase activity: 10 U of this product reacted with 1 μg of λ DNA at 50°C for 16 hours, resulting in no excess DNA bands compared to a 1-hour reaction at 50°C.
4. Enzyme digestion-ligation-re-digestion assay: 10 U of this product was used to digest 1 μg of λ DNA at 16°C with an appropriate amount of T4 DNA ligase. The digested products were then religated and re-cleaved using the same endonuclease.
5. Residual E. coli DNA assay: coli 16s rDNA-specific TaqMan qPCR assay was performed. GenomeResidue ≤ 0.1 pg/ul;
6. Host protein residue: ≤ 50 ppm;
7. Bacterial endotoxin (Endotoxin):LAL-Test, Chinese Pharmacopoeia 2020 Edition, Part IV, Gel Limit Test, General Rules (1143), bacterial endotoxin content ≤ 10 EU/mg;