| Recognition site | RG↑GWCCY
YCCWG↓GR |
| Source | Pseudomonas species PP |
| Assayed on | Lambda DNA (dcm-,HindIII-digest) |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-,HindIII-digest) in 1 hour at 37°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.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 10 - 25 | 10 - 25 | 0 - 10 | 0 - 10 | 100 | 100 |
|
| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store at -20°C. |
| Ligations | After 5-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and more than 80% of these can be recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer Y, BSA |
| Methylation sensitivity | Blocked by overlapping Dcm methylation(CmCWGG): RGGWCCTGG. |
| Inactivation 20 minutes under | 65oC |
| Notes | To obtain 100% activity, BSA should be added
to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
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| MSDS: | Download MSDS as PDF |
| References: | Kileva, E.V., Abdurashitov, M.A., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh. Unpublished observations (1996).
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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
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