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 (2023). A rapid protocol for ribosome profiling of low input samples. Nucleic acids research, 51(13), e68. Oxford University Press 10.1093/nar/gkad459
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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 ∼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: |
Journal Article (Original Article) |
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Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) |
UniBE Contributor: |
Kleemann, Leon, Wu, Jie, Leidel, Sebastian Andreas |
Subjects: |
500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry |
ISSN: |
1362-4962 |
Publisher: |
Oxford University Press |
Language: |
English |
Submitter: |
Pubmed Import |
Date Deposited: |
30 May 2023 09:15 |
Last Modified: |
22 Jul 2023 00:14 |
Publisher DOI: |
10.1093/nar/gkad459 |
PubMed ID: |
37246712 |
BORIS DOI: |
10.48350/183007 |
URI: |
https://boris.unibe.ch/id/eprint/183007 |
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