Do it yourself! - Initial experiences with self-synthesized CsTFA for RNA-SIP analyses

Severin Weis; Susanne Jacksch; Marius Welkerling; Magnus S Schmidt; Markus Egert

doi:10.1016/j.mimet.2022.106432

Do it yourself! - Initial experiences with self-synthesized CsTFA for RNA-SIP analyses

J Microbiol Methods. 2022 Mar:194:106432. doi: 10.1016/j.mimet.2022.106432. Epub 2022 Feb 5.

Authors

Severin Weis¹, Susanne Jacksch¹, Marius Welkerling¹, Magnus S Schmidt², Markus Egert³

Affiliations

¹ Microbiology and Hygiene Group, Institute of Precision Medicine, Faculty of Medical & Life Sciences, Furtwangen University, Villingen-Schwenningen, Germany.
² Organic and Bioorganic Chemistry Lab, Institute of Precision Medicine, Faculty of Medical & Life Sciences, Furtwangen University, Villingen-Schwenningen, Germany.
³ Microbiology and Hygiene Group, Institute of Precision Medicine, Faculty of Medical & Life Sciences, Furtwangen University, Villingen-Schwenningen, Germany. Electronic address: Markus.Egert@hs-furtwangen.de.

PMID: 35134450
DOI: 10.1016/j.mimet.2022.106432

Abstract

Cesium trifluoroacetate (CsTFA) is a gradient medium for isopycnic centrifugation in RNA-based Stable Isotope Probing (RNA-SIP), an important means to link the structure and function of microbial communities. We report a protocol to easily synthesize CsTFA from cesium carbonate (Cs₂CO₃) and trifluoroacetic acid (TFA) and show that self-synthesized CsTFA performs similarly to commercial CsTFA in the separation of isotopically labelled and unlabelled bacterial RNA.

Keywords: Density gradient; Microbial ecology; RNA; Stable Isotope Probing; Ultracentrifugation.

MeSH terms

Carbon Isotopes / chemistry
Centrifugation, Density Gradient / methods
Centrifugation, Isopycnic / methods
Isotope Labeling / methods
Isotopes*
RNA, Bacterial* / genetics
Trifluoroacetic Acid

Substances

Carbon Isotopes
Isotopes
RNA, Bacterial
Trifluoroacetic Acid