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Slama, K.* ; Galliot, A.* ; Weichmann, F.* ; Hertler, J.* ; Feederle, R. ; Meister, G.* ; Helm, M.*

Determination of enrichment factors for modified RNA in MeRIP experiments.

Methods 156, 102-109 (2019)
DOI
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
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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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Merip ; Antibody ; Rna Modification ; M(6)a/n6-methyladenosine ; Tlc; Messenger-rna; Methylation; N-6-methyladenosine; Reveals; Translation; M6a
ISSN (print) / ISBN 1046-2023
e-ISSN 1095-9130
Zeitschrift Methods
Quellenangaben Band: 156, Heft: , Seiten: 102-109 Artikelnummer: , Supplement: ,
Verlag Elsevier
Verlagsort Amsterdam [u.a.]
Begutachtungsstatus Peer reviewed
Institut(e) Monoclonal Antibody (IDO-MAB)