Bpu10 I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain, that carries recombinant plasmids
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); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 50% glycerol. Store at -20°C
Ligations:
After 5-fold overdigestion with enzyme 80% of the DNA fragments can be ligated. Of these 90% can be recut. In the presence of 10% PEG ligation is better.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 1 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer K
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity or incomplete DNA cleavage. We recommend increasing incubation time instead of using an excess of Bpu10I.
The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.
References:
Degtyarev, S.Kh., Zilkin, P.A., Prihodko, G.G., Repin, V.E., Rechkunova, N.I. Mol. Biol. (Mosk) 23: 1051-1056 (1989).
Murat A Abdurashitov, Victor N Tomilov, Valery A Chernukhin, S. Kh. Degtyarev A physical map of human Alu repeats cleavage by restriction endonucleases // BMC Genomics 2008, 9:305
Abdurashitov M.A., Tomilov V.N., Chernukhin V.A., Gonchar D. A., Degtyarev S. Kh. Comparative analysis of human chromosomal DNA digestion with restriction endonucleases in vitro and in silico. // Translated from “Medical genetics” V.6, No 8, pp 29-36, 2007
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006