New England Biolabs, M0308S, T4 PDG (T4 Endonuclease V)

Related Categories DNA Repair Enzymes and Structure-specific Endonucleases Specification Unit Definition One unit is defined as the amount of enzyme that catalyzes the conversion of 0.5 µg of UV irradiated supercoiled pUC19 DNA to > 95% nicked plasmid in a total reaction volume of 20 μl in 30 minutes at 37°C. Nicking is assessed by agarose gel electrophoresis. Irradiated plasmid contains an average of 3-5 pyrimidine dimers. Reaction Conditions 1X T4 PDG Reaction Buffer Supplement with 100 µg/ml Recombinant Albumin, Molecular Biology Grade Incubate at 37°C 1X T4 PDG Reaction Buffer 100 mM NaCl 1 mM DTT 1 mM EDTA 25 mM Na2HPO4 (pH 7.2 @ 25°C) Storage Buffer 10 mM Tris-HCl 250 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 0.15% Triton® X-100 pH 7.4 @ 25°C Heat Inactivation No Unit Assay Conditions 1X T4 PDG Reaction Buffer containing 0.5 µg of UV irradiated supercoiled pUC19 DNA, supplemented with 100 µg/ml Recombinant Albumin in a 20 μl reaction. FAQ Q: What is the activity of T4 PDG in other NEBuffers, including rCutSmart? A: T4 PDG is fully active in rCutSmart and NEBuffer 4 and 75% active in NEBuffers 1-3. All buffers that do not have rAlbumin as a component will need to be supplemented with rAlbumin. Q: What is the molecular weight of T4 PDG? A: The molecular weight of T4 PDG is 16 kDa. Q: Is T4 PDG a tagged protein? A: No. T4 PDG is not a tagged protein. Q: Can the T4 PDG be used in the comet assay? A: We have not tested T4 PDG in the comet assay. Q: Does T4 PDG remove the damaged base? A: Yes. T4 PDG removes the damaged base, unlike UV Damage Endonuclease, which only nicks the DNA. Q: What type of damaged DNA does T4 PDG recognize? A: T4 PDG recognizes cis-syn-cyclobutane pyrimidine dimers caused by UV irradiation.