Nt.BstNB I is a nick endonuclease which cleaves only one strand at the 3’ (N4) of specific sequence as follows :
5’ - - GAGTCNNNN^N - - 3’3’ - - CTCAGNNNNN - - 5’
Source- Recombinant gene from Bacillus stereothermophilus is expressed in E.coli. Application- DNA nicking- DNA recombination Reaction Condition- 1X reaction buffer, incubate at 55°C.10x Reaction Buffer 500 mM Tris-HCl (pH 7.9), 1 M NaCl, 100 mM MgCl₂, 1 mg/ml BSA Storage Buffer- 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.