Endonuclease VIII from E.coli functions as both an N-glycosylase (by excising oxidative base lesions) and an AP lyase (by subsequently cleaving the phosphodiester backbone, leaving terminal phosphates at the 5′ and 3′ ends.). Damaged bases recognized and removed by Endonuclease VIII include urea, 5, 6- dihydroxythymine, thymine glycol, 5-hydroxy-5- methylhydanton, uracil glycol, 6-hydroxy-5, 6-dihydrothymine and methyltartronylurea. Endonuclease VIII also has β and δ lyase activity. Endonuclease VIII is expressed and purified from E.coli.
Applications- Single cell gel electrophoresis (Comet assay)- Alkaline elution- Alkaline unwunding Reagents Supplied & Storage Condition- Endonuclease VIII : 10 unit/㎕, Store at -20°C.- 10x Endonuclease VIII Reaction Buffer : Store at 4°C. Reaction Condition- Endonuclease VIII in 1X Endonuclease VIII Reaction Buffer. Incubate at 37°C. 10x Reaction Buffer- 100 mM Tris-HCl (pH 8.0), 750 mM NaCl, 10 mM DTT Storage Buffer- 10 mM Tris-HCl (pH 8.0), 250 mM NaCl, 0.1 mM EDTA, 50% Glycerol - One unit is defined as the amount of enzyme required to cleave 1 pmol of an oligonucleotide duplex containing a single AP site in 1 hour at 37°C.
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.