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Open Access Green as soon as Postprint is submitted to ZB.
Determination of enrichment factors for modified RNA in MeRIP experiments.
Methods 156, 102-109 (2019)
In the growing field of RNA modification, precipitation techniques using antibodies play an important role. However, little is known about their specificities and protocols are missing to assess their effectiveness. Here we present a method to assess enrichment factors after MeRIP-type pulldown experiments, here exemplified with a commercial antibody against N6-methyladenosine (m(6)A). Testing different pulldown and elution conditions, we measure enrichment factors of 4-5 using m(6)A-containing mRNAs against an unmodified control of identical sequence. Both types of mRNA carry P-32 labels at different nucleotides, allowing their relative quantification in a mixture after digestion to nucleotides, separation by TLC and quantitative phosphorimaging of the labels.
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Scopus SNIP
Web of Science
Times Cited
Times Cited
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3.782
1.039
12
11
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Merip ; Antibody ; Rna Modification ; M(6)a/n6-methyladenosine ; Tlc; Messenger-rna; Methylation; N-6-methyladenosine; Reveals; Translation; M6a
Language
Publication Year
2019
Prepublished in Year
2018
HGF-reported in Year
2018
ISSN (print) / ISBN
1046-2023
e-ISSN
1095-9130
Journal
Methods
Quellenangaben
Volume: 156,
Pages: 102-109
Publisher
Elsevier
Publishing Place
Amsterdam [u.a.]
Reviewing status
Peer reviewed
Institute(s)
CF Monoclonal Antibodies (CF-MAB)
POF-Topic(s)
30201 - Metabolic Health
Research field(s)
Helmholtz Diabetes Center
PSP Element(s)
G-502210-001
WOS ID
WOS:000462417800014
Scopus ID
85056191525
PubMed ID
30394295
Erfassungsdatum
2018-11-07