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Updated: Nov 3, 2025
Selective Capture of 5-hydroxymethylcytosine from Genomic DNA
Published on: October 5, 2012
Quantitative and Single-Nucleotide Resolution Profiling of RNA 5-Methylcytosine
Jun Li1, Xingyu Wu1, Trung Do1
1Department of Molecular and Biomedical Sciences, School of Biological Sciences, The University of Adelaide, Adelaide, SA, Australia.
Abstract:
RNA has coevolved with numerous posttranscriptional modifications to sculpt interactions with proteins and other molecules. One of these modifications is 5-methylcytosine (m5C) and mapping the position and quantifying the level in different types of cellular RNAs and tissues is an important objective in the field of epitranscriptomics. Both in plants and animals bisulfite conversion has long been the gold standard for detection of m5C in DNA but it can also be applied to RNA. Here, we detail methods for highly reproducible bisulfite treatment of RNA, efficient locus-specific PCR amplification, detection of candidate sites by sequencing on the Illumina MiSeq platform, and bioinformatic calling of non-converted sites.
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