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A rapid protocol for ribosome profiling of low input samples
Meindl, Andreas, Romberger, Markus, Lehmann, Gerhard, Eichner, Norbert, Kleemann, Leon
, Wu, Jie, Danner, Johannes, Boesl, Maria, Mesitov, Mikhail, Meister, Gunter, König, Julian, Leidel, Sebastian Andreas und Medenbach, Jan
(2023)
A rapid protocol for ribosome profiling of low input samples.
Nucleic Acids Research 51 (13), e68-e68.
Veröffentlichungsdatum dieses Volltextes: 04 Sep 2023 13:59
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.54612
Zusammenfassung
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 ...
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.
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| Dokumentenart | Artikel | ||||
| Titel eines Journals oder einer Zeitschrift | Nucleic Acids Research | ||||
| Verlag: | OXFORD UNIV PRESS | ||||
|---|---|---|---|---|---|
| Ort der Veröffentlichung: | OXFORD | ||||
| Band: | 51 | ||||
| Nummer des Zeitschriftenheftes oder des Kapitels: | 13 | ||||
| Seitenbereich: | e68-e68 | ||||
| Datum | 29 Mai 2023 | ||||
| Institutionen | Biologie und Vorklinische Medizin > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I | ||||
| Identifikationsnummer |
| ||||
| Stichwörter / Keywords | IN-VIVO; TRANSLATION; REVEALS; INITIATION; DYNAMICS; SITES; CELLS; | ||||
| Dewey-Dezimal-Klassifikation | 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie | ||||
| Status | Veröffentlicht | ||||
| Begutachtet | Ja, diese Version wurde begutachtet | ||||
| An der Universität Regensburg entstanden | Ja | ||||
| URN der UB Regensburg | urn:nbn:de:bvb:355-epub-546123 | ||||
| Dokumenten-ID | 54612 |
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