Description
Recognition site and hydrolysis position:
RAATTY | R↑AATTY |
YTTAAR | YTTAA↓R |
Source: Arthrobacter citreus
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 50°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W + BSA
Enzyme activity (%):
B | G | O | W | Y | ROSE |
25 | 50 | 50 | 100 | 10 | 100 |
Storage conditions: 20 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 10 mM 2-mercaptoethanol; 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 20 u.a. of enzyme for 16 hours at 50°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Degtyarev, S.Kh., Kolyhalov, A.A., Rechkunova, N.I., Abdurashitov, M.A., Nucleic Acids Res. 20: 3789 (1992).
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