Endonuclease V, a repair enzyme found in E. coli, is a 3'-endonuclease involved in DNA repair, which initiates removal of deaminated bases from damaged DNA. Endonuclease V recognizes DNA containing deoxyinosines, a deamination product of deoxyadenosine, on double-stranded DNA, single-stranded DNA containing abasic sites (ap) or urea, base mismatches, insertion/deletion mismatches, hairpin or unpaired loops, flaps and pseudo-Y structures. Endonuclease V requires accessory proteins to remove the damaged base. Endonuclease V is expressed and purified from E.coli.
Applications- High-throughput methods for mutation research- Studies in mutagenesis and DNA repair- Cleavage of oligonucleotides containing deoxyinosines and a weaker affinity for oligonucleotides containing base mismatches Reagents Supplied & Storage Condition- Endonuclease V : 10 unit/㎕, Store at -20°C.- 10x Endonuclease V Reaction Buffer : Store at 4°C. Reaction Condition- Endonuclease V in 1X Endonuclease V Reaction Buffer. Incubate at 37°C. 10x Reaction Buffer- 500 mM Potassium Acetate, 200 mM Tris-acetate, 100 mM Magnesium Acetate, 10 mM DTT Storage Buffer- 10 mM Tris-HCl (pH 8.0), 300 mM NaCl, 1 mM DTT, 1 mM EDTA, 50% Glycerol, 0.15% Triton X-100 - One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34 mer oligonucleotide duplex containing a single deoxyinosine site in a total reaction volume of 10 μl in 1 hour at 37°C.
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.