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Research Paper

Isolation and Posttranscriptional Modification Analysis of Native BC1 RNA from Mouse Brain

Timofey S. Rozhdestvensky, Pamela F. Crain and Juergen Brosius

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We present a simple and general affinity method, based on size fractionation and nucleic acid complementarity, to isolate sufficient amounts of native RNA molecules for further physicochemical studies, such as modification state of nucleotides. In the case presented here, we purified four micrograms of dendritic neuronal BC1 RNA from 130 grams of mouse brain (initially yielding a total of 200 mg RNA). Directly combined liquid chromatography-electrospray ionization mass spectrometry (LC/MS) analysis revealed no base or sugar backbone modifications in native BC1 RNA, despite earlier indications that C-54 could be methylated in vitro (Cm5, position 54).

Authors

Timofey S. Rozhdestvensky

University of Münster, Münster, Germany

Pamela F. Crain

Unviersity of Utah, Salt Lake City, Utah

Juergen Brosius

Institute of Experimental Pathology, ZMBE, University of Münster; Münster, Germany

This is an open-access article

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If the document does not open, please right-click on the link (control-click on a Macintosh) and select the option to save the file to disk.