PspL I

Enzyme name PspL I
Prototype SplI
Known analogs BsiWI, Pfl23II, SplI, SunI
SKU SE-E223
Turbo version Not available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… C▼GTACG …3'
3'… GCATG▲C …5'
Source Pseudomonas species L
Optimal buffer SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
25502525100100
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA (HindIII-digest)
Storage conditions 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 2-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer Y, BSA
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.
References
  1. Unpublished data Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh Unpublished observations (1995)
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