| Recognition site | ↑GATC
CTAG↓ |
| Source | Bacillus stearothermophillus MB |
| Assayed on | Lambda DNA (dam-) |
| 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. |
| Optimal SE-buffer | O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 10 - 25 | 25 - 50 | 100 | 75 - 100 | 10 - 25 | 100 |
|
| Optimal temperature | 65oC |
| 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 |
10 X SE-buffer O |
| 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’ |
| Inactivation 20 minutes under | 80oC |
| MSDS: | Download MSDS as PDF |
| References: | Kovalenko, K., Kileva, E.V., Mezentseva, N.V., Mikhnenkova, N.A., Dedkov, V.S., Degtyarev, S.K. Unpublished observations. (2004)
|
Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
|
