Description
Recognition site and hydrolysis position:
GATC | ↑GATC |
CTAG | CTAG↓ |
Source: Bacillus stearothermophillus MB
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 65°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dam-)
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 25 | 100 | 75 | 10 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 200 μg/ml BSA. Store at -20°C.
Ligations:
After 5-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 65°C
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dam methylation(GmATC): GATC.
Not blocked by CG methylation.
Not cut hemimethylated site:5`- G(6mA)TC-3` / 3`-CTAG-5`
Notes:
References:
Kovalenko, K., Kileva, E.V., Mezentseva, N.V., Mikhnenkova, N.A., Dedkov, V.S., Degtyarev, S.K. Unpublished observations. (2004)