BstKT I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus KT
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dam-)
Optimal SE-buffer: SE-buffer W
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 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 5 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer W
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dam methylation (G^mATC) GATC.
Not blocked by CG methylation.
Cut hemimethylated site:5`- GmATC-3` / 5`-GATC-3`
Notes:
References:
Nadeev A.N., Chernukhin V. A., Sevastyanova O.O., Tomilova J.E., Shinkarenko N.M., Evdokimov A.A., Degtyarev S. Kh. BstKTI, a New dam-Sensitive Neoschizomer of Restriction Endonuclease MboI, which is Able to Cleave Hemimethylated Substrate.// Biotekhnologia (Moscow), No.2, 5-10 (2006). (In Russian)