Bse8 I

Description

Recognition site and hydrolysis position:

Source: Bacillus species 8
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
Long incubation is not recommended. Extended incubations and high concentration of the enzyme may result in star activity. Reagents Supplied with Enzyme 10 X SE-buffer G
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dam-methylation (GmATC): 5’-GmATCN^NNATC-3’/3’-CTmAGN^NNTAG.
Blocked by overlapping (5mC)-methylation: 5’-GATNN^NNAT(5mC)-3’/3’-(5mC)TANN^NNTAG-5’.
Cuts hemimethylated sequence: 5’-GATNN^NNAT(5mC)-3’/3’-CTANN^NNTAG-5’.

Notes: High enzyme concentration (more than 5-fold overdigestion on 1 μg substrate) may result in star activity.
References:
Repin, V.E., Serov, G.D., Puchkova, L.I., Tereshenko, T.A., Lebedev, L.R., Chigikov, V.E. Bioorg. Khim. 19: 583-585 (1993).