T4 Endonuclease V

Applications\nStudy of UV damage to DNA and its repair, including DNA damage in single cells. Detection of differential UV damage repair of transcribed sequences. Detection of UV mutational hotspots.\nT4 Endonuclease V functions as part of a base excision repair pathway to recognize and remove\ncyclobutane pyrimidine dimers. The enzyme binds to UV-irradiated DNA and processively\nscans the DNA until a pyrimidine dimer is encountered. T4 Endonuclease V then cleaves\nthe glycosyl bond of the 5'-pyrimidine of the dimer and the 3' phosphodiester bond, resulting\nin breakage of the DNA strand.1,2 Strand breakage is evident during subsequent electrophoresis\nas a change in conformation (i.e., from covalent closed circular to relaxed circular molecules), or as the appearance of unique DNA fragments. The enzyme is a small protein that does not require divalent cations or other cofactors.1\n\nSource:\ngene denV in E. coli\n\nActivity:\n20u/ul\n\nUnit Definition:\nOne unit will change 1ug of UV-irradiated plasmid DNA from covalently closed circles (form I) to nicked closed circles (form II) in 30 minutes at 37 degrees C.\n\nActivity Assay: \nThe activity assay is performed in a 50ul reaction containing 50mM Tris-HCl pH 7.5) and 5mM using plasmid DNA irradiated for 1 minute at 254nm.\n\nContaminating Activity Assays: \nT4 Endonuclease V is free of detectable exonuclease, non-T4 Endonuclease V-specific endonuclease and RNase activities as judged by electrophoresis after incubation of 1mg of various DNA substrates with 500-1000U of enzyme at 37 degrees C for 16 hours.\n\nStorage Buffer: \n50 mM Tris-HCl, pH 7.5, 100mM sodium chloride, 0.1mM EDTA, 1mM DTT, 0.1% Triton X-100, 50% glycerol.\n\nWorking Solution:\nDilute the enzyme in 0.01M EDTA with 0.1mg/ml nuclease-free BSA, and buffer of choice (0.04M sodium phosphate or equivalant). The presence of EDTA is critical.\n\nQuality Control: \nT4 Endonuclease V is free of detectable nonspecific exo-and endonuclease activities and RNase activity.\n\nStorage and Stability:\nStable for one year at -20C.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

T0200

Size

100U

Form

Supplied as a frozen solution in 0.01M phosphate buffer, pH 6.5 containing 0.01M EDTA, 0.001M DTT and 20% glycerol.

References

1. Friedberg, E.C. et al., (1995) DNA Repair and Mutagenesis, ASM Press, Washington D.C., 164.\n2. Schrock, R.D. and Lloyd, R.S. (1993) J. Biol. Chem. 268, 880.

Alternative Names

gene denV in E. coli