Source

- Nocardia otitidis-caviarum.

Reaction Conditions
- Buffer 3 + BSA. 50 mM Tris-HCl, pH 7.9, 100 mM NaCl, 10 mM MgCl₂, 1 mM DTT. Supplement with 100 ㎍/ml BSA. Incubate at 37℃.

Concentration & Storage Condition
- Store at -20℃. 10 unit/㎕ in 20 mM Tris-HCl, pH 7.4, 200 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 200 ㎍/ml BSA, 50% (v/v) glycerol, 0.15% Triton X-100.

Heat Inactivation Condition
- 65℃ for 20 min.

Enzyme Dilution
- Dilute with Storage Buffer.

QC Tests
- Activity, ligation and recutting, exo and endonuclease activity test, SDS-PAGE purity, performance tests.

Methylase Sensitivity

- Blocked by CpG methylation (cleavage of mammalian genomic DNA is blocked). 

 

Note

- Supercoiled plasmids may required up to 5 folds more Not I for complete digestion than linear DNAs.