Ssp I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain, that carries the cloned gene SspI from Sphaerotilus species
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 K
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 90% 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 10 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer K, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by methylation A^mATATT.
Notes: High enzyme concentration may result in star activity.
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:
Schildkraut, I., Grandoni, R. unpublished observations.
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