Restriction Enzymes

 

NotI     NotI is a restriction enzyme which recognizes the following sequence: GC▼GGCC GC CG CCGG▲CG Concentration   10u/μl   Reaction condition Recommended Buffer: Buffer O+ 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA . Incubation temperature: 37°C Thermal inactivation: 65°C   Source   Nocardia otitdis-caviarum.   Frequency of Cutting   λ Ad-2 ΦX174 pUC18 M13mp18 pBR322 0 7 0 0 0 0   Storage   -20°C   Storage buffer   20mM Tris-HCl (pH 7.8 at 25°C), 100mM NaCl, 0.1mM EDTA, 10mM 2-mercaptoethanol, 0.02% Triton X-100, 0.2mg/ml BSA and 50% glycerol   Ligation and Recleavage   After 50-fold overdigestion with NotI, approximately more than 90% of the DNA fragments can be ligated and recut   Methylation Effect   NotI cuts neither GCGGm5CCGC nor GCGGCm5CGC, but cuts GCGGCCGm5C Blocked by CG methylation   Notes Assayed on Adenovirus-2 DNA     Order   Back