Bsa29 I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus 29
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 G + BSA
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
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 G, BSA (except E205T and E206T).
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dam methylation (G^mATC): GATCGATC.
Blocked by CG methylation.
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:
Repin, V.E., Degtyarev, S.K. Unpublished observations.