Ajn I

Enzyme nameAjn I
PrototypeEcoRII
Known analogsEcoRII, Psp6 I, PspGI, BptI, BseBI, Bst2U I, BstNI, BstOI, MvaI
SKUSE-E473
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'… ▼CCWGG …3'
3'… GGWCC▲ …5'
SourceAcinetobacter johnsonii R2
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 temperature55 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
2510102510050
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 3-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 6 u.a. of enzyme for 16 hours at 55°C.
Methylation sensitivityNot blocked by dcm methylation CCWGG
Reagents supplied with enzyme10 X SE-buffer Y
NotesAt 37°C activity is about 10% from maximum.
References
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