BspAC I

Description

Recognition site and hydrolysis position:

Source: Bacillus species AC
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer O + BSA
Enzyme activity (%):

Storage conditions: 10 mM KH₂PO₄(pH 7.2); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme more than 95% of the Lambda DNA fragments can be ligated with T4 DNA Ligase at 16°C and 50% of these can be recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer O, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by CG methylation.
Notes: To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
BspACI has a non-palindromic recognition site.
References:
Chernukhin, V.A., Tomilova, J.E., Dedkov, V.S., Janobilova, Z.K.. Unpublished observations(2006).
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006