EcoICR I

Enzyme nameEcoICR I
PrototypeSacI
Known analogsEcl136II, Psp124B I, SacI, SstI
SKUSE-E469
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'… GAG▼CTC …3'
3'… CTC▲GAG …5'
SourceEscherichia coli ICR
Optimal bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
7510000755
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onLambda DNA (HindIII-digest)
Storage conditions10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
LigationAfter 5-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 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer G, BSA
NotesTo obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml Do not use BSA for long incubation.
References
  1. External publication Sugisaki H., Maekawa Y., Kanazawa S., Takanami M. Bull. Inst. Chem. Res. Kyoto Univ. 60: 328-335 (1982).
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