Absolute Quantification of Noncoding RNA by Microscale Thermophoresis

Jacob, Dominik; Thüring, Kathrin; Galliot, Aurellia; Marchand, Virginie; Galvanin, Adeline; Ciftci, Akif; Scharmann, Karin; Stock, Michael; Roignant, Jean-Yves; Leidel, Sebastian Andreas; Motorin, Yuri; Schaffrath, Raffael; Klassen, Roland; Helm, Mark (2019). Absolute Quantification of Noncoding RNA by Microscale Thermophoresis. Angewandte Chemie (International ed.), 58(28), pp. 9565-9569. Wiley-VCH 10.1002/anie.201814377

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Accurate quantification of the copy numbers of noncoding RNA has recently emerged as an urgent problem, with impact on fields such as RNA modification research, tissue differentiation, and others. Herein, we present a hybridization-based approach that uses microscale thermophoresis (MST) as a very fast and highly precise readout to quantify, for example, single tRNA species with a turnaround time of about one hour. We developed MST to quantify the effect of tRNA toxins and of heat stress and RNA modification on single tRNA species. A comparative analysis also revealed significant differences to RNA-Seq-based quantification approaches, strongly suggesting a bias due to tRNA modifications in the latter. Further applications include the quantification of rRNA as well as of polyA levels in cellular RNA.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Leidel, Sebastian Andreas

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1433-7851

Publisher:

Wiley-VCH

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

07 Jan 2020 15:22

Last Modified:

07 Jan 2020 15:22

Publisher DOI:

10.1002/anie.201814377

BORIS DOI:

10.7892/boris.137058

URI:

https://boris.unibe.ch/id/eprint/137058

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