Bsp19 I

Enzyme name Bsp19 I
Prototype NcoI
Known analogs NcoI
SKU SE-E047
Turbo version Available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… C▼CATGG …3'
3'… GGTAC▲C …5'
Source Bacillus species 19
Optimal buffer SE-buffer 2W (20 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 200 mM NaCl; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
0105075105
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
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.
Nonspecific hydrolysis 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.
Methylation sensitivity Bsp19I cuts hemimethylated site 5`-(5mC)CATGG-3`/3`-GGTACC-5` and doesn't cut methylatedsites 5`-(5mC)CATGG-3`/3`-GGTAC(5mC)-5` and 5`-(4mC)CATGG-3`/3`-GGTAC(4mC)-5`.
Reagents supplied with enzyme 10 X SE-buffer 2W, BSA
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
  2. Unpublished data Repin, V.E., Rechkunova, N.I., Degtyarev, S.Kh Unpublished observations.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).
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