| Name | | Bsp19 I |   |
| | Cat. # | E047 | E048 | E047T | E048T | | Package, u.a. | 1000 | 5000 | 1000 | 5000 | | Concentration, u.a./ml | 10000-20000 | 10000-20000 | 10000-20000 | 10000-20000 |
| 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. | | 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 37°C.
| | Reagents Supplied with Enzyme |
10 X SE-buffer 2W, BSA (except E047T and E048T).
For E047T and E048T an additional 10X SE-buffer ROSE is supplied.
| | 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 Unpublished observations.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
| 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
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