Sph I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned Sph I gene from Streptomyces phaeochromogenes
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 G
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme more than 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 G, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
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:
Fuchs, L.Y., Covarrubias, L., Escalante, L., Sanchez, S., Bolivar, F. Gene 10:39-46 (1980).