CciN I

Enzyme name CciN I
Prototype NotI
Known analogs NotI
SKU SE-E203
Turbo version Available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… GC▼GGCCGC …3'
3'… CGCCGG▲CG …5'
Source Curtobacterium citreus N
Optimal buffer SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
25507575100100
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Adenovirus-2 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Adenovirus-2 DNA
Storage conditions 10 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.
Ligation After 5-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of DNA Ad-2 with 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity Blocked by CG methylation.
Reagents supplied with enzyme 10 X SE-buffer Y.
Notes High enzyme concentration may result in star activity.
References
  1. V.S. Dedkov, N.I. Rechkunova, E.A. Prihodko, E.V. Kileva, Y.S. Kusner, V.A. Verchozina, S.Kh. Degtyarev CciNI, an isoschisomer of NotI from Curtobacterium citreum recognizes 5′-GC^GGCCGC-3′. // Gene, No. 157, 99-100 (1995).
  3. V.E. Repin, S.Kh. Degtyarev, N.N. Karpyshev, V.E. Chizhikov Nucleic acid fragment and recombinant plasmid DNA pDKR 85 for testing of rare cutters restriction endonucleases SfiI and NotI. // Soviet Union patent SU 1433020 (1986). (In Russian)
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