Journal article
Absolute Quantification of Noncoding RNA by Microscale Thermophoresis



Publication Details
Authors:
Schaffrath, R.
Publisher:
WILEY-V C H VERLAG GMBH
Publication year:
2019
Journal:
Angewandte Chemie International Edition
Pages range:
9565-9569
Volume number:
58
Issue number:
28
Start page:
9565
End page:
9569
Number of pages:
5
ISSN:
1433-7851

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.


Keywords
hybridization, microscale thermophoresis, RNA quantification, tRNA stability

Last updated on 2021-19-01 at 15:35