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A Fluorescent Reverse-Transcription Assay to Detect Chemical Adducts on RNA.

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Herein, we detail a novel reverse-transcription (RT) assay to directly detect chemical adducts on RNA. We optimize a fluorescence quenching assay to detect RT polymerization and employ our approach to… Click to show full abstract

Herein, we detail a novel reverse-transcription (RT) assay to directly detect chemical adducts on RNA. We optimize a fluorescence quenching assay to detect RT polymerization and employ our approach to detect N1-alkylation of inosine, an important post-transcriptional modification, using a phenylacrylamide as a model compound. We anticipate our approach can be expanded to identify novel reagents that form adducts with RNA and further explored to understand the relationship between RT processivity and natural post-transcriptional modifications in RNA.

Keywords: assay detect; adducts rna; detect chemical; transcription assay; chemical adducts; reverse transcription

Journal Title: Biochemistry
Year Published: 2022

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