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A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA by Weiling Maggie Cai; Yok Hian Chionh; Fabian Hia; Chen Gu; Stefanie Kellner; Megan E. McBee; Chee Sheng Ng; Yan Ling Joy Pang; Erin G. Prestwich; Kok Seong Lim; I. Ramesh Babu; Thomas J. Begley; Peter C. Dedon is a book available to read on EtoBox.

What is A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA about?

Here we describe an analytical platform for systems-level quantitative analysis of modified ribonucleosides in any RNA species, with a focus on stress-induced reprogramming of tRNA as part of a system of translational control of cell stress response. This chapter emphasizes strategies and caveats for each of the seven steps of the platform workflow: (1) RNA isolation, (2) RNA purification, (3) RNA hydrolysis to individual ribonucleosides, (4) chromatographic resolution of ribonucleosides, (5) identification of the full set of modified ribonucleosides, (6) mass spectrometric quantification of ribonucleosides, (6) interrogation of ribonucleoside datasets, and ( 7) mapping the location of stresssensitive modifications in individual tRNA molecules. We have focused on the critical determinants of analytical sensitivity, specificity, precision, and accuracy in an effort to ensure the most biologically meaningful data on mechanisms of translational control of cell stress response. The methods described here should find wide use in virtually any analysis involving RNA modifications.

Author

Weiling Maggie Cai; Yok Hian Chionh; Fabian Hia; Chen Gu; Stefanie Kellner; Megan E. McBee; Chee Sheng Ng; Yan Ling Joy Pang; Erin G. Prestwich; Kok Seong Lim; I. Ramesh Babu; Thomas J. Begley; Peter C. Dedon

Published

2015

Language

EN