PspPP I

Description

Recognition site and hydrolysis position:

Source: Pseudomonas species PP
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.
Assayed on:
Lambda DNA/HindIII
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

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
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dcm methylation(C^mCWGG): RGGWCCTGG.
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.
References:
Kileva, E.V., Abdurashitov, M.A., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh. Unpublished observations (1996).