Bpm I

Description

Recognition site and hydrolysis position:

Source: Bacillus pumilus
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
Optimal SE-buffer: SE-buffer W + BSA
Enzyme activity (%):

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 2-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and 95% may be recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer W, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dcm-methylation (CmCWGG):
5′- C(5mC)TGGAG(N)₁₆-3′
3′- GGA(5mC)CTC(N)₁₄-5′
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:
Degtyarev, S.Kh., Morgan P. Unpublished observation (1992).