Biomaterial Database

A filter elution assay for the simultaneous detection of DNA double and single strand breaks. 1996

J Lu, and M R Kaeck, and C Jiang, and G Garcia, and H J Thompson

Division of Laboratory Research, AMC Cancer Research Center, Denver, Colorado 80214, USA.

This paper reports a gravity-flow filter elution assay that permits the simultaneous detection of DNA double strand breaks (DSBs, as neutral elutable DNA) and DNA single strand breaks (SSBs, as alkaline elutable DNA). This assay is based on a modification of a conventional gravity-flow alkaline elution procedure. The elution fraction resulting from cell lysis before the application of the alkaline elution buffer provides an estimate of DNA DSBs, while the fraction eluted by the alkaline buffer (pH 12.3) measures the extent of DNA SSBs. Data show that, in addition to its good detection sensitivity for SSBs, this method provides a sensitive measure of DNA DSBs. As an illustration of its specificity, this method unambiguously distinguished DNA SSBs induced by X-irradiation of mouse leukemia L1210 cells from DNA DSBs occurring in cells undergoing nucleosomal DNA fragmentation and apoptosis. The neutral elution fraction is a good predictor of loss of cell viability induced either by selenite at pharmacological levels or by growth factor depletion. The filter elution method reported is more specific and informative than a nick translation assay for the detection of DNA strand breaks.

UI	MeSH Term	Description	Entries
D007939	Leukemia L1210	An experimental LYMPHOCYTIC LEUKEMIA of mice.	Leukemia L 1210,L 1210, Leukemia,L1210, Leukemia
D009690	Nucleic Acid Conformation	The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.	DNA Conformation,RNA Conformation,Conformation, DNA,Conformation, Nucleic Acid,Conformation, RNA,Conformations, DNA,Conformations, Nucleic Acid,Conformations, RNA,DNA Conformations,Nucleic Acid Conformations,RNA Conformations
D004247	DNA	A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).	DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004249	DNA Damage	Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS.	DNA Injury,DNA Lesion,DNA Lesions,Genotoxic Stress,Stress, Genotoxic,Injury, DNA,DNA Injuries
D005374	Filtration	A process of separating particulate matter from a fluid, such as air or a liquid, by passing the fluid carrier through a medium that will not pass the particulates. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Filtrations
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D051379	Mice	The common name for the genus Mus.	Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D053938	DNA Fragmentation	Splitting the DNA into shorter pieces by endonucleolytic DNA CLEAVAGE at multiple sites. It includes the internucleosomal DNA fragmentation, which along with chromatin condensation, are considered to be the hallmarks of APOPTOSIS.	DNA Degradation, Apoptotic,Apoptotic DNA Degradation,Fragmentation, DNA
D018038	Sodium Selenite	The disodium salt of selenious acid. It is used therapeutically to supply the trace element selenium and is prepared by the reaction of SELENIUM DIOXIDE with SODIUM HYDROXIDE.	Disodium Selenite,Monosodium Selenite,Selenous Acid Disodium Salt,Selenous Acid, Disodium Salt,Sodium Biselenite,Sodium Selenite Pentahydrate,Biselenite, Sodium,Pentahydrate, Sodium Selenite,Selenite Pentahydrate, Sodium,Selenite, Disodium,Selenite, Monosodium,Selenite, Sodium