|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 (%)|
|10 - 25||10 - 25||0 - 10||0 - 10||100||100|
|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.
|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 Unpublished observations (1996). |
|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