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Absolute Quantification of Noncoding RNA by Microscale Thermophoresis.

ABSTRACT: 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.

SUBMITTER: Jacob D

PROVIDER: S-EPMC6617968 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Absolute Quantification of Noncoding RNA by Microscale Thermophoresis.

Jacob Dominik D Thüring Kathrin K Galliot Aurellia A Marchand Virginie V Galvanin Adeline A Ciftci Akif A Scharmann Karin K Stock Michael M Roignant Jean-Yves JY Leidel Sebastian A SA Motorin Yuri Y Schaffrath Raffael R Klassen Roland R Helm Mark M

Angewandte Chemie (International ed. in English) 20190502 28

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 ...[more]

PMID: 30892798

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