| Species | Bacillus stearothermophilus |
| Synonyms | BsaI restriction endonuclease |
| Expression System | E.coli |
| Molecular Weight | 64.9kDa (Reducing) |
| Purity | >95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 10 mM Tris-HCl、300 mM NaCl、1 mM DTT、0.1 mM EDTA、500 µg/ml BSA、50% Glycerol （pH 7.4 @ 25°C） |
| Stability & Storage | Store at -25 ~ -15℃for 2 years |
| Reference |
[1]Zhu Z, Samuelson J C, Zhou J, et al.Engineering Strand-specific DNA NickingEnzymes from the Type IIS Restriction Endonucleases BsaI, BsmBI, andBsmAI[J].Journal of Molecular Biology, 2004,337(3):573-583. [2] Lee J H, Won H J, Oh E S,etal. Golden GateCloning-Compatible DNA Replicon/2A-Mediated Polycistronic Vectors forPlants[J]. Frontiers in Plant Science, 2020, 11. |
BsaIis a Type IIs restriction enzyme that can recognize non-palindromic sequencesand cut outside of the recognition sequence. It is commonly used for GoldenGate assembly and enzymatic cleavage of plasmids to prepare linear DNAfragments with poly (A/T/G/C) endings and obtain specific sticky ends.
Recognitionsite:
5'-GGTCTC(N)1↓-3'
3'-CCAGAG(N)5↑-5'
StorageSolution: 20 U/μl BsaⅠ、10 mM Tris-HCl、300 mM NaCl、1 mM DTT、0.1 mM EDTA、500 µg/ml BSA、50% Glycerol（pH 7.4 @ 25°C）
10*Reaction Buffer：500 mM Potassium Acetate、200 mM Tris-acetate、100 mM Magnesium Acetate、1mg/ml BSA（pH 7.4 @ 25°C）
Thisstep is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled upaccording to experimental needs.
1）Add the following components insequence
| Components | Volume |
Plasmid DNA |
1μg DNA |
10*Reaction Buffer |
5μl |
BsaⅠ (20 U/μl) |
1μl |
RNase-free ddH2O |
Up to 50μl |
2）Incubate at 37°C 1h
3）DNA linearization iscomplete, and subsequent experiments can be performed.
1. star activity may result from a glycerol concentration of >5%; 2. Please avoid repeated freeze-thaw cycles.
The results of 1μg pUC57 plasmid digestion separated under different quantity of BsaⅠ, The reaction was incubated for 60 minutes at 37°C, and 1% agarose gel was used for electrophoresis analysis after reaction.
M, marker;
Lane 1 1μg pUC57;
Lane 2 1μg pUC57 add 1U BsaⅠ
Lane 3 1μg pUC57 add 2U BsaⅠ
1μg (R: reducing
condition, N: non-reducing condition).
