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.
CITATION STYLE
Amort, T., Sun, X., Khokhlova-Cubberley, D., & Lusse, A. (2017). Transcriptome-wide detection of 5-methylcytosine by bisulfite sequencing. In Methods in Molecular Biology (Vol. 1562, pp. 123–142). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6807-7_9
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