Description
Recognition site and hydrolysis position:
CCTNNNNNAGG | CCTNN↑NNNAGG |
GGANNNNNTCC | GGANNN↓NNTCC |
Source: Bacillus stearothermophillus EM
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
50 | 50 | 25 | 25 | 100 | 100 |
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 5-fold overdigestion with enzyme about 60% of the DNA fragments can be ligated and recut. Of these 90% 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 65°C
Reagents Supplied with Enzyme:
Methylation sensitivity: Not blocked by overlapping Dcm methylation(CmCWGG): CCTGGNNNAGG,
CCTNNNCCAGG.
Notes:
References:
Abdurashitov, M.A., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2000).