| 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 | R | | 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 |
| Ligations | 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.
|
| 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Unpublished observations (2001).
|
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
|
