Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 58, Issue 28, Pages 9565-9569Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201814377
Keywords
fluorescence; hybridization; microscale thermophoresis; RNA quantification; tRNA stability
Categories
Funding
- DFG within the SPP1784 [HE3397/8-1, HE3397/13-2, SCHA750/20-2, KL2937/1-2, RO4681/6-1, LE3260/2-1]
- ANR-DFG grant HTRNA-Mod [ANR-13-ISV8-0001]
- Lorraine Region (France)
- IMB Core Facility
- Agence Nationale de la Recherche (ANR) [ANR-13-ISV8-0001] Funding Source: Agence Nationale de la Recherche (ANR)
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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.
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