Product 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 at 37°C 1 h; 3. DNA linearization is complete, and subsequent experiments can be performed. Notes 1. Heat inactivation condition: incubate at 80°C for 20min. 2. Please operate with lab coats and disposable gloves,for your safety. Ver.EN20230725