T4 Endonuclease V, also known as PDG (pyrimidine dimer glycosylase), has N-glycosylase and apurinic/apyrimidinic lyase (AP lyase) activities. T4 Endonuclease V locates and binds cis-syn-cyclobutane pyrimidine dimers caused by UV irradiation. The enzyme cleaves the glycosyl bond of the 5´ end of the pyrimidine dimer and breaks the phosphodiester bond at the AP site. T4 Endonuclease V is expressed and purified from E.coli.
Applications- Study of UV damage to DNA and its repair- Single cell gel electrophoresis (Comet assay)- Detection of UV mutational hotspots Reagents Supplied & Storage Condition- T4 Endonuclease V : 10 unit/㎕, Store at -20°C- 10x T4 Endonuclease V Reaction Buffer : Store at 4°C- 10 mg/ml BSA : Sore at. 4°C Reaction Condition- T4 Endonuclease V in 1X T4 Endonuclease Reaction Buffer and 0.1 mg/ml BSA. Incubate at 37°C. 10x Reaction Buffer- 1 M NaCl, 10 mM DTT, 10 mM EDTA, 250 mM Na2HPO4 (pH 7.2) Storage Buffer- 10 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM DTT, 0.1 mM EDTA, 50% Glycerol Unit Definition- One unit of T4 Endonuclease V converts 1 µg of UV-irradiated plasmid DNA from covalently closed circles to nicked form in 30 minutes at 37°C. QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.