RT Journal Article
SR Electronic
T1 A rapid protocol for ribosome profiling of low input samples
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.09.23.509038
DO 10.1101/2022.09.23.509038
A1 Andreas Meindl
A1 Markus Romberger
A1 Gerhard Lehmann
A1 Norbert Eichner
A1 Leon Kleemann
A1 Jie Wu
A1 Johannes Danner
A1 Maria Boesl
A1 Mikhail Mesitov
A1 Gunter Meister
A1 Julian König
A1 Sebastian Leidel
A1 Jan Medenbach
YR 2022
UL http://biorxiv.org/content/early/2022/09/23/2022.09.23.509038.abstract
AB Ribosome profiling provides quantitative, comprehensive, and high-resolution snapshots of cellular translation by the high-throughput sequencing of short mRNA fragments that are protected from nucleolytic digestion by ribosomes. 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 RNA. 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.Competing Interest StatementJM is a consultant to siTOOLs Biotech GmbH, Martinsried, Germany