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METHODS OF DETECTING DNA N-GLYCOSYLASES, METHODS OF DETERMINING N-GLYCOSYLASE ACTIVITY, AND N-GLYCOLSYLASE ASSAY KITS
METHODS OF DETECTING DNA N-GLYCOSYLASES, METHODS OF DETERMINING N-GLYCOSYLASE ACTIVITY, AND N-GLYCOLSYLASE ASSAY KITS
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机译:检测DNA N-糖基化酶的方法,N-糖基化酶活性的测定方法和N-糖基化酶测定试剂盒
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摘要
The invention includes methods of detecting glycosylases. A test sample is mixed with substrate polynucleotide. A primer and a polymerase are added. An endonuclease is provided and a probe oligonucleotide sequence labeled with first and second labels is utilized for detection. The invention includes N-glycosylase assay methods. A test sample is mixed with substrate polynucleotide and formation of an abasic site is detected by forming a product that is complementary to a portion of the substrate sequence ending at the abasic site. The product is dissociated and is extended utilizing a polymerase. A probe is hybridized to the product and is cleaved. The invention includes synthetic substrates, transcription primers and probe molecules. The invention also includes an N-glycosylase detection kit including a substrate polynucleotide, an endonuclease and a dual-labeled probe having a fluorescent label and a quencher moiety.
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