Nt.Fok I is a nick endonuclease which cleaves only one strand at the 3’ (N9) of specific sequence as follows.
This enzyme is engineered by chimeric fusion of the restriction domain of Nt.BstNB I and the DNA recognition part of Fok I.5’ - - GGATGN(9) ^ NNNN - - 3’3’ - - CCTACN(9)NNNN - - 5’
Source- Recombinant fusion gene of DNA recognition domain of Fok I and the cleavage domain of Nt.BstNB I is expressed in E.coli. Application- DNA nickingReaction Condition1X reaction buffer, incubate at 37°C. 10x Reaction Buffer- 100 mM Tris-HCl (pH 7.9), 500 mM NaCl, 100 mM MgCl₂, 10 mM DTT - 10 mM Tris-HCl (pH 7.4), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 ㎍/ml BSA, 50% Glycerol
Unit Definition- One unit is defined as the amount of enzyme required to convert 1 µg of supercoiled DNA to open circular form in 1 hour at 37°C in a total reaction volume of 50 µl. Heat Inactivation- 80°C for 20 min QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.