Acc36 I

Enzyme nameAcc36 I
PrototypeBspMI
Known analogsBfuAI, BspMI, BveI
SKUSE-E289
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IIEiType IIE enzymes interact with two recognition sites: one site is cleaved, while the second site acts as an allosteric effector site., Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… ACCTGC(N)4▼ …3'
3'… TGGACG(N)8▲ …5'
SourceAcinetobacter calcoaceticus 36
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
25255050100100
Unit definitionOne 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 onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 100 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 enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 4 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y
NotesHigh enzyme concentration may result in star activity
References
  1. Unpublished data Gonchar, D.A., Shinkarenko, N.M., Dedkov, V.S., Degtyarev, S.Kh Unpublished observations. 1999
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