Description
Recognition site and hydrolysis position:
| GACNNNNGTC | GACNN↑NNGTC |
| CTGNNNNCAG | CTGNN↓NNCAG |
Source: Bacillus stearothermophillus PA
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 | 25 | 50 | 50 | 100 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 200 μg/ml BSA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme less than 5% of the DNA fragments can be ligated
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 25°C
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration and incubation at 65°C for 16 hours results in star activity.
References:
Tomilova, J.E., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.Kh. Unpublished observations (2000).
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