Direkt zum Inhalt

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 ; Medenbach, Jan

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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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftNucleic Acids Research
Verlag:OXFORD UNIV PRESS
Ort der Veröffentlichung:OXFORD
Band:51
Nummer des Zeitschriftenheftes oder des Kapitels:13
Seitenbereich:e68-e68
Datum29 Mai 2023
InstitutionenBiologie und Vorklinische Medizin > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I
Identifikationsnummer
WertTyp
10.1093/nar/gkad459DOI
Stichwörter / KeywordsIN-VIVO; TRANSLATION; REVEALS; INITIATION; DYNAMICS; SITES; CELLS;
Dewey-Dezimal-Klassifikation500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
StatusVeröffentlicht
BegutachtetJa, diese Version wurde begutachtet
An der Universität Regensburg entstandenJa
URN der UB Regensburgurn:nbn:de:bvb:355-epub-546123
Dokumenten-ID54612

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