Recognition site | GCG↑C C↓GCG |
Source | Bacillus stearothermophilus HH |
Assayed on | Lambda DNA |
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. |
Optimal SE-buffer | Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA |
Enzyme activity (%) | B | G | O | W | Y | Rose | 75 - 100 | 50 - 75 | 25 - 50 | 50 - 75 | 100 | 100 |
|
Optimal temperature | 50oC |
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 |
Ligation | After 40-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 Enzyme |
10 X SE-buffer Y, BSA |
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. |
Inactivation 20 minutes | No |
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.
|
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Shapovalova, M.A., Abdurashitov, M.A., Tumanova, I.Y., Dedkov, V.S., Popichenko, D.V., Degtyarev, S.K.
Unpublished observations (2001).
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