T4 PDG (T4 Endonuclease V)Product information
- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
Product SourcePurified from an E. coli strain carrying a plasmid encoding T4 denV gene.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|T4 PDG Reaction Buffer||10X|
|Purified BSA||-20||10 mg/ml|
Advantages and Features
- DNA damage studies
- Single cell gel electrophoresis (comet assay)
Properties and Usage
Unit DefinitionOne 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.
1X T4 PDG Reaction Buffer
Supplement with 100X Purified BSA
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
10 mM Tris-HCl
100 mM NaCl
0.1 mM DTT
0.1 mM EDTA
pH 7.5 @ 25°C
Unit Assay Conditions1X T4 PDG Reaction Buffer containing 0.5 µg of UV irradiated supercoiled pUC19 DNA, supplemented with 100 µg/ml BSA in a 20 µl reaction.
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
- Exonuclease Activity (Radioactivity Release):
The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
- Non-Specific DNase Activity (16 hour):
The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
- Protein Purity (SDS-PAGE):
The physical purity is assessed by comparing contaminating protein bands in a concentrated sample to the protein of interest band in a sample of known dilution. The purity is determined by SDS-PAGE.
- For best results incubation time should be 30 minutes or less.
- Addition of 1 µl of phenol to the sample before loading will improve electrophoresis by stripping the protein from the DNA.
- Warm buffer to room temperature as it precipitates at 4°C.
- Higgins, K.L. and Lloyd, R.S. (1987). Mutation Research. 183, 117-121.