PspN4 I

Description

Recognition site and hydrolysis position:

Source: Pseudomonas species N4
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by
5`-GGNN(5mC)C-3`/3-C(5mC)NNGG-5` or
5`-GGNN(5mC)C-3`/3-CCNNGG-5`methylation
Notes:
References:
Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh. Unpublished observations (1995).
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 4, pp 19-27, 2007