Biomaterial Database

32P-postlabelling analysis of bulky aromatic adducts. 1993

R C Gupta

Preventive Medicine and Environmental Health, University of Kentucky, Lexington 40506.

The 32P-postlabelling assay has emerged as a major tool for detecting DNA adducts induced by structurally diverse carcinogens, particularly bulky aromatics. The assay comprises enzymatic degradation of DNA to 3'-mononucleotides, enrichment of adducts, 5'-32P-labelling, adduct separation by TLC, and detection and quantitation of adducts. This report describes (1) an improved solvent extraction procedure for isolating DNA from tissues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios issues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios.

UI	MeSH Term	Description	Entries
D008722	Methods	A series of steps taken in order to conduct research.	Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D010761	Phosphorus Radioisotopes	Unstable isotopes of phosphorus that decay or disintegrate emitting radiation. P atoms with atomic weights 28-34 except 31 are radioactive phosphorus isotopes.	Radioisotopes, Phosphorus
D002274	Carcinogens, Environmental	Carcinogenic substances that are found in the environment.	Environmental Carcinogens
D002855	Chromatography, Thin Layer	Chromatography on thin layers of adsorbents rather than in columns. The adsorbent can be alumina, silica gel, silicates, charcoals, or cellulose. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Chromatography, Thin-Layer,Thin Layer Chromatography,Chromatographies, Thin Layer,Chromatographies, Thin-Layer,Thin Layer Chromatographies,Thin-Layer Chromatographies,Thin-Layer Chromatography
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
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012997	Solvents	Liquids that dissolve other substances (solutes), generally solids, without any change in chemical composition, as, water containing sugar. (Grant & Hackh's Chemical Dictionary, 5th ed)	Solvent