Quantitative and Single-Nucleotide Resolution Profiling of RNA 5-Methylcytosine
| Autor: | Xingyu Wu, Pei Qin Ng, Jun Li, Trung Do, Jing Zhao, Iain R. Searle, Vy Nguyen, Alice Burgess, Rakesh David |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0303 health sciences Bisulfite sequencing RNA Computational biology 01 natural sciences DNA sequencing law.invention Bisulfite 03 medical and health sciences 5-Methylcytosine chemistry.chemical_compound chemistry law Epitranscriptomics Polymerase chain reaction DNA 030304 developmental biology 010606 plant biology & botany |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071613733 |
| DOI: | 10.1007/978-1-0716-1374-0_9 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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