Acl I

Enzyme nameAcl I
PrototypeAclI
Known analogsPsp1406I
SKUSE-E011
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'… AA▼CGTT …3'
3'… TTGC▲AA …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
000010080
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; 200 μg/ml BSA; 0.05% Triton X-100; 50% glycerol; Store at -20°C.
LigationAfter 2-fold overdigestion with enzyme 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 2 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by CG methylation
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesTo 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
  1. S.Kh. Degtyarev, M.A. Abdurashitov, A.A. Kolyhalov, N.I. Rechkunova Acl I, a new restriction endonuclease from Acinetobacter calcoaceticus recognizing 5′-AA^CGTT -3′. // Nucleic Acids Research, Vol. 20, No. 14, 37-87 (1992).
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