Process for directly determining apurinic and apyrimidinic...

Details Process for directly determining apurinic and apyrimidinic... Process for directly determining apurinic and apyrimidinic...

C - Chemistry, Metallurgy

12

Q

150/8.5

C12Q 1/68 (2006.01)

Patent

CA 1212023

A b s t r a c t A simple and fast process is described which serves to directly determine apurinic and apyrimidinic sites (AP sites) in DNA using [14C] methoxyamine which, after loss of a purine or pyrimidine base, respectively, reacts with the resultant alde- hyde groups of the deoxyribose groups. The incorporation of the [14C] methoxyamine in DNA is propro- tional to the number of AP sites. Since methoxyamine does not cause the DNA to degrade, the unreacted AP sites can be mea- sured in order to determine the radioactivity of the acid- insoluble fraction. The process lends itself to the analysis of DNA-damages such as those which are physically or chemically produced. Moreover it is suitable to demonstrate and determine the activity of DNA glycosylases, in particular of uracil-DNA glycosylases.

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