Description
This product is a type IIS restriction endonuclease derived from the recombinant protein encoded by the BsaI gene in Bacillus sphaericus expressed by E.coli. Its recognition sequence is 5'-GGTCTCN1/N5-3'. Use to digest plasmids to prepare poly(A/T/G/C)-terminated linearized DNA fragments to obtain specific cohesive ends.
This product is produced in accordance with GMP process requirements and provided in a liquid form.
Specifications
|
Expression Host |
Recombinant E. coli with BasI gene |
|
Reaction Temperature |
37℃ |
|
Storage Buffer |
10 mM Tris-HCl, 0.2 M NaCl, 0.1 mM EDTA, 1mM DTT, 50% Glycerol |
|
Unit Definition |
1 unit: The amount of enzyme required to digest 1 μg of substrate DNA within 1 h at 37℃ in a 50 μL system. |
|
Application |
1.Digest the plasmid to prepare a linearized DNA fragment at the end of Poly (A/T/C/G); 2.Digestion of DNA to obtain specific sticky ends； 3.Linearize plasmid template before in-vitro transcription. |
Components
|
Components No. |
Name |
10661ES03 (1 KU) |
10661ES10 (10 KU) |
10661ES60 (100 KU) |
|
10661 |
BsaI GMP-grade (20 U/μL) |
50 μL |
500 μL |
5 mL |
Storage
This product should be stored at -25 ~ -15℃ for two years.
Instructions
Experimental methods
50 μL reaction system
This step is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled up according to experimental needs.
- 1.Add the following components in sequence:
|
Components |
Volume |
|
Plasmid DNA |
1-2 μg |
|
10×Digestion Buffer 4 |
5.0 μL |
|
BsaI (20 U/μL) |
1.0 μL |
|
RNase-free ddH2O |
Up to 50 μL |
【Note】10× Digestion Buffer 4(Cat#10668ES): 500 mM Potassium Acetate，200 mM Tris-acetate，100 mM Magnesium Acetate，1 mg/ml OsrHSA, pH7.9@25℃
- 2.Incubate at37°C 1 h;
- 3.DNA linearization is complete, and subsequent experiments can be performed.
Notes
- Heat inactivation condition: incubate at80°C for20min.
- Please operate with lab coats and disposable gloves，for your safety.