14-C METHOXYAMINE - A METHOD FOR REACTION OF APURINIC - APYRIMIDINIC SITES WITH THE QUANTITATIVE ASSAY OF AP SITES IN DNA
A simple and rapid method is described for the determination of AP (apurinic/apyrimidinic) sites in DNA. The method involves the reaction of 14-C-methoxyamine with the aldehyde group present in the deoxyribose moiety after a base loss. Studies with alkylated - depurinated DNA and with uracil containing polydeoxyribonucleotides depyrimidinated by uracil-DNA glycosylase show that methoxyamine reacts with both apurinic and apyrimidinic sites in a rapid and exhaustive way. Under standard conditions (30-min incubation with 5 mM methoxyamine at 37 degrees C, pH 7.2) untreated DNA is almost unreactive and the 14-C-methoxyamine incorporation in DNA is proportional to the number of AP sites. Since the methoxyamine reaction is free from any degradative effect on DNA, AP sites may be estimated from a simple determination of the acid insoluble radioactivity.
Bibliographic Reference: BIOCHIMICA ET BIOPHYSICA ACTA, VOL. 740 (1983), PP. 410-416
Record Number: 1989122025000 / Last updated on: 1987-01-01
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