Chapter title |
Transcriptome-Wide Detection of 5-Methylcytosine by Bisulfite Sequencing
|
---|---|
Chapter number | 9 |
Book title |
RNA Methylation
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6807-7_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6805-3, 978-1-4939-6807-7
|
Authors |
Amort, Thomas, Sun, Xueguang, Khokhlova-Cubberley, Daria, Lusser, Alexandra, Thomas Amort, Xueguang Sun, Daria Khokhlova-Cubberley, Alexandra Lusser |
Editors |
Alexandra Lusser |
Abstract |
While low-throughput RNA bisulfite sequencing is the method of choice to assess the methylation status of specific cytosines in candidate RNAs, the combination of bisulfite treatment of RNA with today's high-throughput sequencing techniques opens the door to methylation studies at nucleotide resolution on a transcriptome-wide scale. Below we describe a protocol for the transcriptome-wide analysis of total or fractionated poly(A)RNA in cells and tissues. Although the nature of the bisulfite sequencing protocol makes it comparably easy to translate from a low to a high-throughput approach, several critical points require attention before starting such a project. We describe a step-by-step protocol for planning and performing the experiment and analyzing the data. |
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