Abstract
RNA modification mapping by mass spectrometry (MS) is based on the use of specific ribonucleases (RNases) that generate short oligonucleotide digestion products which are further separated by nano-liquid chromatography and analyzed by MS and MS/MS. Recent developments in MS instrumentation allow the possibility to deeply explore posttranscriptional modifications. Notably, development of nano-liquid chromatography and nano-electrospray drastically increases the detection sensitivity and allows the identification and sequencing of RNA digested fragments separated and extracted from two-dimensional polyacrylamide gels, as long as the mapping and characterization of ribonucleotide modifications.
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Acknowledgments
The authors would like to thank Gérard Keith for the total E. coli tRNA preparation and Dominique Burnouf for critical reading of the manuscript and useful comments. We are grateful to Pascale Romby and Eric Westhof for their constant support. This work was supported by Labex NetRNA.
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Antoine, L., Wolff, P. (2020). Mapping of Posttranscriptional tRNA Modifications by Two-Dimensional Gel Electrophoresis Mass Spectrometry. In: Arluison, V., Wien, F. (eds) RNA Spectroscopy. Methods in Molecular Biology, vol 2113. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0278-2_8
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DOI: https://doi.org/10.1007/978-1-0716-0278-2_8
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