Aco I

Description

Recognition site and hydrolysis position:

Source: Acinetobacter calcoaceticus
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA(dam-, dcm-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 3-fold overdigestion with AcoI more than 90% of Lambda DNA fragments can be ligated with T4 DNA Ligase at 16°C and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of AcoI for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping dcm methylation (C^mCWGG): CCTGGCCR.
Notes:
References:
Chernukhin, V.A., Belichenko, O.A., Tomilova, J.E., Dedkov, V.S., Doroganov, A.O.. Unpublished observations. (2006)