| Recognition site | CCCAG↑C
G↓GGTCG |
| Source | Geobacillus stearothermophilus Y |
| Assayed on | Lambda DNA |
| 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. |
| Optimal SE-buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 10 - 25 | 25 - 50 | 75 - 100 | 100 | 75 - 100 | 100 |
|
| Optimal temperature | 70oC |
| 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 |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes | No |
| 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.
|
| MSDS: | Download MSDS as PDF |
| 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)
|
Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
|
