BsaI restriction endonuclease HYJ110
BsaI restriction endonuclease is a mutant protein encoded by a gene from Bacillus stearothermophilus expressed recombinantly in Escherichia coli. It is a class IIS restriction endonuclease that recognizes specific sites with the same specificity as the native enzyme but with reduced star activity. Its recognition sequence and cleavage site are as follows:
5'······GGTCTC(N)···············3'
3'······CCAGAG(NNNNN)······5
Components
|
Components |
1KU |
5KU |
20KU |
200KU |
2000KU |
|
BsaI (20 U/μL) |
50 uL |
250 uL |
1 mL |
10 mL |
100 mL |
|
10×BsaI 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 1 year (avoid repeated freezing and thawing).
Storage buffer
10 mM Tris-HCl, 200 mM NaCl, 1 mM DTT 0.1 mM EDTA, 200 µg/ml Recombinant Albumin,
50% Glycerol. (pH 7.4 @ 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 37°C within 1 hour.
Quality control
1. Protein purity (SDS-PAGE): BsaI purity was ≥95% as determined by SDS-PAGE analysis.
2. RNase activity:20 U of this product and 1.6 μg of MS2 RNA were reacted at 37°C for 16 hours, resulting in no change in the RNA electrophoretic band.
3. Nonspecific DNase activity: 20 U of this product and 1 μg of PhiX174 DNA were reacted at 37°C for 16 hours, resulting in no excess DNA electrophoretic bands.
4. Enzyme digestion-ligation-re-digestion assay: 1 μg of λDNA/HindIII was digested with 20 U of this product at 16°C using an appropriate amount of T4 DNA ligase. The digested products can be religated. After the ligated products are recovered, they can be re-cleaved using the same endonuclease.
5. E.coli DNA Residual Test: 2 U of nucleic acid remaining in this product was detected by TaqMan qPCR specific for E. coli 16s rDNA, and the E. coli genome remained at ≤1 copy.
6. Endotoxin: LAL-Test, Chinese Pharmacopoeia 2020 Edition, Part IV, Gel Limit Test, General Rules (1143), bacterial endotoxin content ≤10 EU/mg