Gsa I

Description

Recognition site and hydrolysis position:

Source: Geobacillus stearothermophilus Y
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 70°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme about 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of DNA with 20 u.a. of enzyme for 16 hours at 70°C. Reagents Supplied with Enzyme 10 X SE-buffer W, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: Do not use BSA for long incubation.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
References:
Dedkov V.S., Mikhnenkova N.A., Kileva E.V., Tarasova G.V., Gonchar D.A., Akishev A.G., Chernukhin V.A., Shkirja S.S., Degtyarev S.Kh.
New restriction endonuclease GsaI, neoschizomer of BseY I, recognizes 5’-CCCAGC(-1/-5)-3’.// Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov (Moscow), V.4, No.1, p.19-24 (2008). (In Russian)