Product info: Bsp19 I Name Bsp19 I      Cat. # E047 E048 Package, u.a. 1000 5000 Concentration, u.a./ml 20000 20000 Recognition site C↑CATGG GGTAC↓C Source Bacillus species 19 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 37°C in a total reaction volume of 50 μl. Optimal SE-buffer 2W (20 mM Tris-HCl (pH 8,5 at 25°C); 10 mM MgCl2; 200 mM NaCl; 1 mM DTT) + BSA Enzyme activity (%) B G O W Y Rose 0 - 10 10 - 25 50 - 75 75 - 100 10 - 25 5 Optimal temperature 37oC Storage conditions 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C. Ligation After 20-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 40 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer 2W, BSA Methylation sensitivity Bsp19I cuts hemimethylated site 5`-(5mC)CATGG-3`/3`-GGTACC-5` and doesn't cut methylated sites 5`-(5mC)CATGG-3`/3`-GGTAC(5mC)-5` and 5`-(4mC)CATGG-3`/3`-GGTAC(4mC)-5`. Inactivation 20 minutes under 65oC 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. Quality control Restriction Endonucleases: Quality Control MSDS: Download MSDS as PDF References: Repin, V.E., Rechkunova, N.I., Degtyarev, S.Kh. Unpublished observations. Repin, V.E., Lebedev, L.R., Andreeva, I.S., Puchkova, L.I., Zernov, Y.P., Serov, G.D., Tereshchenko, T.A., Aphinogenova, G.N., Pustoshilova, N.M. Biotekhnologiya 0: 18-27 (1998).  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 // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007