title: Nucleotide-Level Profiling of m⁵C RNA Methylation.
abstract: Mapping the position and quantifying the level of 5-methylcytosine (m(5)C) as a modification in different types of cellular RNA is an important objective in the emerging field of epitranscriptomics. Bisulfite conversion has long been the gold standard for detection of m(5)C in DNA but it can also be applied to RNA. Here, we detail methods for bisulfite treatment of RNA, locus-specific PCR amplification and detection of candidate sites by sequencing on the Illumina MiSeq platform.
authors.affiliation: Genome Biology Department, The John Curtin School of Medical Research (JCSMR), The Australian National University, Acton, Canberra, ACT, Australia.
authors.forename: Tennille
authors.initials: T
authors.lastname: Sibbritt
authors.affiliation: Genome Biology Department, The John Curtin School of Medical Research (JCSMR), The Australian National University, Acton, Canberra, ACT, Australia.
authors.forename: Andrew
authors.initials: A
authors.lastname: Shafik
authors.affiliation: Genomics and Epigenetics Department, Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW, Australia.
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Clark
authors.affiliation: Genome Biology Department, The John Curtin School of Medical Research (JCSMR), The Australian National University, Acton, Canberra, ACT, Australia. thomas.preiss@anu.edu.au.|Victor Chang Cardiac Research Institute, Darlinghurst, Sydney, NSW, Australia. thomas.preiss@anu.edu.au.
authors.forename: Thomas
authors.initials: T
authors.lastname: Preiss
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism*; D019175:DNA Methylation / Q000235:genetics; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D009711:Nucleotides / Q000235:genetics; D012313:RNA / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*
chemical_list: D009711:Nucleotides; D012313:RNA; D044503:5-Methylcytosine
keywords: 5-Methylcytosine; Bisulfite conversion; Epitranscriptomics; MiSeq; Next-generation sequencing