Recognition site | RG↑GWCCY YCCWG↓GR |
Source | Pseudomonas species PP |
Assayed on | Lambda DNA/HindIII |
Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA/HindIII 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 | Rose | 10 - 25 | 10 - 25 | 0 - 10 | 0 - 10 | 100 | 100 |
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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. |
Ligation | 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.
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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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Quality control | Restriction Endonucleases: Quality Control |
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 Unpublished observations (1996).
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