Slama, Kaouthar and Galliot, Aurellia and Weichmann, Franziska and Hertler, Jasmin and Feederle, Regina and Meister, Gunter and Helm, Mark (2019) Determination of enrichment factors for modified RNA in MeRIP experiments. METHODS, 156. pp. 102-109. ISSN 1046-2023, 1095-9130
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MESSENGER-RNA; METHYLATION; N-6-METHYLADENOSINE; REVEALS; TRANSLATION; M6A; MeRIP; Antibody; RNA modification; M(6)A/N6-methyladenosine; TLC |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 16 Apr 2020 06:50 |
| Last Modified: | 16 Apr 2020 06:50 |
| URI: | https://pred.uni-regensburg.de/id/eprint/27428 |
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