Development of a chemiluminescent reagent that recognizes guanine in nucleic acid, and its use for chemiluminescence detection of DNA

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A novel reagent, 3', 4', 5'-trimethoxyphenylglyoxal has been found to specifically convert guanine bases of a nucleic acid in a neutral or alkaline phosphate solution to the corresponding chemiluminescent derivatives. The chemiluminogenic derivatization technique offers a facile method of the sensitive imaging detection for a DNA probe tailing with deoxyriboguanylic acid oligomers. A cooled charge-coupled device camera is usable to detect visually the chemiluminescence emitted from the derivatized probe that was hybridized with a target DNA fragment on a Nylon membrane. The proposed method permits the quantitative detection of the probe covalently attached with a 5'- (G) _<15>TT (G) _<15>TT deoxyoligomer at 3' terminus, indicating a detection limit of 0.1 pmol on the membrane. The detection sensitivity can be increased with larger contents of deoxyriboguanylic acid in the probe molecule.