Target-Specific Profiling of RNA m5C Methylation Level Using Amplicon Sequencing

Tennille Sibbritt; Ulrike Schumann; Andrew Shafik; Marco Guarnacci; Susan J. Clark; Thomas Preiss

doi:10.1007/978-1-0716-1851-6_21

Target-Specific Profiling of RNA m⁵C Methylation Level Using Amplicon Sequencing

Tennille Sibbritt, Ulrike Schumann, Andrew Shafik, Marco Guarnacci, Susan J. Clark, Thomas Preiss^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

1 Citation (Scopus)

Abstract

Mapping the position and quantifying the level of 5-methylcytosine (m⁵C) as a modification in different types of cellular RNA is an important objective in the field of epitranscriptomics. Bisulfite conversion has long been the gold standard for the detection of m⁵C 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.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	375-392
Number of pages	18
DOIs	https://doi.org/10.1007/978-1-0716-1851-6_21
Publication status	Published - 26 Oct 2021

Publication series

Name	Methods in Molecular Biology
Volume	2404
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

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10.1007/978-1-0716-1851-6_21

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title = "Target-Specific Profiling of RNA m5C Methylation Level Using Amplicon Sequencing",

abstract = "Mapping the position and quantifying the level of 5-methylcytosine (m5C) as a modification in different types of cellular RNA is an important objective in the field of epitranscriptomics. Bisulfite conversion has long been the gold standard for the detection of m5C 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.",

keywords = "5-methylcytosine, Bisulfite conversion, Epitranscriptomics, MiSeq, Next-generation sequencing",

author = "Tennille Sibbritt and Ulrike Schumann and Andrew Shafik and Marco Guarnacci and Clark, {Susan J.} and Thomas Preiss",

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T1 - Target-Specific Profiling of RNA m5C Methylation Level Using Amplicon Sequencing

AU - Sibbritt, Tennille

AU - Schumann, Ulrike

AU - Shafik, Andrew

AU - Guarnacci, Marco

AU - Clark, Susan J.

AU - Preiss, Thomas

PY - 2021/10/26

Y1 - 2021/10/26

N2 - Mapping the position and quantifying the level of 5-methylcytosine (m5C) as a modification in different types of cellular RNA is an important objective in the field of epitranscriptomics. Bisulfite conversion has long been the gold standard for the detection of m5C 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.

AB - Mapping the position and quantifying the level of 5-methylcytosine (m5C) as a modification in different types of cellular RNA is an important objective in the field of epitranscriptomics. Bisulfite conversion has long been the gold standard for the detection of m5C 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.

KW - 5-methylcytosine

KW - Bisulfite conversion

KW - Epitranscriptomics

KW - MiSeq

KW - Next-generation sequencing

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U2 - 10.1007/978-1-0716-1851-6_21

DO - 10.1007/978-1-0716-1851-6_21

M3 - Chapter

T3 - Methods in Molecular Biology

SP - 375

EP - 392

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Target-Specific Profiling of RNA m⁵C Methylation Level Using Amplicon Sequencing

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