DNA adducts occur through environmental, therapeutic, dietary, oxygen stress, and aging processes. A modified thin-layer chromatographic (TLC) technique can assess base composition and adduct formation. This requires labeling DNA by "shot-gun" 5'-phosphorylation of representative 32P-α-deoxyribonucleotide monophosphates. Subsequent 3'-monophosphate digest "sister exchanges" a radioactive 32PO4 2- to the neighboring cold nucleotide. Separation in two-dimensional polyethyleneimine-cellulose TLC is carried out in acetic acid, (NH4)2SO4, and (NH4)HSO4. The technique was applied to control DNA, cold substition of dUMP, methylation, depurination, and pBR322. This technique quantifies low-molecular-mass adducts and DNA integrity both in vivo and in vitro.
|Original language||English (US)|
|Number of pages||15|
|Journal||Journal of Chromatography B: Biomedical Sciences and Applications|
|State||Published - Feb 7 1992|
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