Description
Recognition site and hydrolysis position:
| GCGC | GCG↑C |
| CGCG | C↓GCG |
Source: Bacillus stearothermophilus HH
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 50°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y + BSA
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 75 | 50 | 25 | 50 | 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 50-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of DNA with 100 u.a. of enzyme for 16 hours at 65°C.Reagents Supplied with Enzyme10 X SE-buffer Y, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by
5`-G(5mC)GC-3`/3-CG(5mC)G-5` or
5`-G(5mC)GC-3`/3`-CGCG-5`methylation.
Not blocked by
5`-GCG(5mC)-3`/3`-(5mC)GCG-5` or
5`-GCG(5mC)-3`/3`-CGCG-5`methylation.
Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml
Do not use BSA for long incubation.
References:
Shapovalova, M.A., Abdurashitov, M.A., Tumanova, I.Y., Dedkov, V.S., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).
