Aco I

Enzyme nameAco I
PrototypeCfrI
Known analogsCfrI, EaeI
SKUSE-E499
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… Y▼GGCCR …3'
3'… RCCGG▲Y …5'
SourceAcinetobacter calcoaceticus
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
50755025100100
Unit definitionOne 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 onLambda DNA
Storage conditions10 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.
LigationAfter 3-fold overdigestion with AcoI more than 90% of Lambda DNA fragments can be ligated with T4 DNA Ligase at 16°C and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of AcoI for 16 hours at 37°C.
Methylation sensitivityBlocked by overlapping dcm methylation (C m CWGG) CC TGG CCR.
Reagents supplied with enzyme10 X SE-buffer Y
References
  1. Unpublished data Chernukhin, V.A., Belichenko, O.A., Tomilova, J.E., Dedkov, V.S., Doroganov, A.O Unpublished observations. (2006)
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