Titlebook: Epitranscriptomics; Methods and Protocol Narendra Wajapeyee,Romi Gupta Book 2019 Springer Science+Business Media, LLC, part of Springer Nat

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Book 2019 targeted and unbiased high-throughput analysis associated with post-transcriptional RNA modification. The chapters in this book also cover specific topics such as transcriptome-wide detection of 5-methylcytosine; HAMR; iRNA-2OM; genome-wide annotation of circRNAs; immune-northern blotting; and dete

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Bisulfite Sequencing of RNA for Transcriptome-Wide Detection of 5-Methylcytosine, aspects need to be considered before starting such a project. Below we describe a detailed step-by-step protocol for planning and performing a transcriptome-wide bisulfite sequencing experiment and subsequent data analysis to determine methyl-cytosine in poly(A)RNA from cells and tissues.

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,Direct Chemical Biotinylation of RNA 5′-Ends Using a Diazo Reagent,n-reagent, as exemplified on a 110 nucleotide RNA obtained via in vitro transcription. The method exploits the fact that, under neutral buffer conditions (~pH 6.8), the 5′-phosphate carries the only mildly acidic proton in the RNA molecule, which allows for selective functionalization at that site using diazo reagents.

Subjugate

Identification of Methylated Transcripts Using the TRIBE Approach,ion. Here, we describe a complementary technique of standard meRIP-seq/miCLIP-seq approaches to identify methylated RNA using a?low amount of?material. We believe this approach can be applied in vivo to identify methylated targets in specific tissues or subpopulations of cells.

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Peter Kopietz,Lorenz Bartosch,Florian SchützPR/Cas9-based approach. This strategy is?relatively simple approach to track?. at different stages of cell differentiation, providing mechanistic insights into the initiation, maintenance, and establishment of X inactivation.