DNA precipitation assay: a rapid and simple method for detecting DNA damage in mammalian cells
When mammalian cells are lysed in 2% sodium dodecyl sulphate detergent followed by addition of an equal volume of 0.12 M potassium chloride, a precipitate forms that can be collected by low-speed centrifugation. This precipitate contains the cell protein and nucleic acid in close association with protein. In the absence of DNA damage, most of the DNA precipitates, but when DNA strand breaks are created by exposing cells to ionizing radiation or toxic chemicals, DNA is released from the protein and remains in the supernatant after centrifugation. The proportion of DNA remaining in the supernatant is thus a measure of the amount of DNA damage. This assay is characterized in terms of optimum cell number and pH and dose-response curves for DNA damage and cell survival following ionizing radiation, MNNG, BCNU, and VP-16 are shown. Sensitivity, simplicity, speed, and large sample handling capacity should allow wide application of this new assay to a variety of questions concerning DNA damage and repair.
- Research Organization:
- British Columbia Cancer Research Centre, Vancouver (Canada)
- OSTI ID:
- 7138915
- Journal Information:
- Environ. Mol. Mutagen.; (United States), Vol. 11:4
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA
GENETIC RADIATION EFFECTS
STRAND BREAKS
BIOASSAY
RADIOINDUCTION
XENOBIOTICS
GENETICS
DOSE-RESPONSE RELATIONSHIPS
FIBROBLASTS
HAMSTERS
IONIZING RADIATIONS
MUTAGENS
PRECIPITATION
SURVIVAL TIME
ULTRACENTRIFUGATION
ANIMAL CELLS
ANIMALS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGY
CENTRIFUGATION
CONNECTIVE TISSUE CELLS
GENETIC EFFECTS
MAMMALS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATION EFFECTS
RADIATIONS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture
560300 - Chemicals Metabolism & Toxicology