BstC8 I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus C8
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 55°C. Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by GC methylation: 5`-G(5mC)NNGC-3`/3`-CGNN(5mC)G-5`.
Notes: At 37°C activity is 50% from maximum.
References:
Abdurashitov, M.A., Dedkov, V.S., Mikhnenkova, N.A., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).
Dedkov, V.S. Novel M.BstC8I Methyltransferase Forms 5′-Gm5CNNGC-3′.Investigation of Restriction Endonuclease Sensitivity to M.BstC8I Methylation.// Mol. Gen., Microbiol. and Virology, Vol. 27. No.1. P. 40-47 (2012).