Meindl, Andreas and Romberger, Markus and Lehmann, Gerhard and Eichner, Norbert and Kleemann, Leon and Wu, Jie and Danner, Johannes and Boesl, Maria and Mesitov, Mikhail and Meister, Gunter and Koenig, Julian and Leidel, Sebastian Andreas and Medenbach, Jan (2023) A rapid protocol for ribosome profiling of low input samples. NUCLEIC ACIDS RESEARCH, 51 (13). ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
Ribosome profiling provides quantitative, comprehensive, and high-resolution snapshots of cellular translation by the high-throughput sequencing of short mRNA fragments that are protected by ribosomes from nucleolytic digestion. While the overall principle is simple, the workflow of ribosome profiling experiments is complex and challenging, and typically requires large amounts of sample, limiting its broad applicability. Here, we present a new protocol for ultra-rapid ribosome profiling from low-input samples. It features a robust strategy for sequencing library preparation within one day that employs solid phase purification of reaction intermediates, allowing to reduce the input to as little as 0.1 pmol of similar to 30 nt RNA fragments. Hence, it is particularly suited for the analyses of small samples or targeted ribosome profiling. Its high sensitivity and its ease of implementation will foster the generation of higher quality data from small samples, which opens new opportunities in applying ribosome profiling.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | IN-VIVO; TRANSLATION; REVEALS; INITIATION; DYNAMICS; SITES; CELLS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 07 Mar 2024 13:50 |
| Last Modified: | 07 Mar 2024 13:50 |
| URI: | https://pred.uni-regensburg.de/id/eprint/58916 |
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