Gutenberg Open Science: Absolute and relative quantification of RNA modifications via biosynthetic isotopomers

Please use this identifier to cite or link to this item: http://doi.org/10.25358/openscience-8002

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DC Field	Value	Language
dc.contributor.author	Kellner, Stefanie	-
dc.contributor.author	Ochel, Antonia	-
dc.contributor.author	Thüring, Kathrin	-
dc.contributor.author	Spenkuch, Felix	-
dc.contributor.author	Neuman, Jennifer	-
dc.contributor.author	Sharma, Sunny	-
dc.contributor.author	Entian, Karl-Dieter	-
dc.contributor.author	Schneider, Dirk	-
dc.contributor.author	Helm, Mark	-
dc.date.accessioned	2022-10-14T07:24:30Z	-
dc.date.available	2022-10-14T07:24:30Z	-
dc.date.issued	2014
dc.identifier.uri	https://openscience.ub.uni-mainz.de/handle/20.500.12030/8017	-
dc.description.abstract	In the resurging field of RNA modifications, quantification is a bottleneck blocking many exciting avenues. With currently over 150 known nucleoside alterations, detection and quantification methods must encompass multiple modifications for a comprehensive profile. LC MS/MS approaches offer a perspective for comprehensive parallel quantification of all the various modifications found in total RNA of a given organism. By feeding <sup>13</sup> C-glucose as sole carbon source, we have generated a stable isotope-labeled internal standard (SIL-IS) for bacterial RNA, which facilitates relative comparison of all modifications. While conventional SIL-IS approaches require the chemical synthesis of single modifications in weighable quantities, this SIL-IS consists of a nucleoside mixture covering all detectable RNA modifications of Escherichia coli, yet in small and initially unknown quantities. For absolute in addition to relative quantification, those quantities were determined by a combination of external calibration and sample spiking of the biosynthetic SIL-IS. For each nucleoside, we thus obtained a very robust relative response factor, which permits direct conversion of the MS signal to absolute amounts of substance. The application of the validated SIL-IS allowed highly precise quantification with standard deviations <2% during a 12-week period, and a linear dynamic range that was extended by two orders of magnitude.	en_GB
dc.description.sponsorship	DFG, Open Access-Publizieren Universität Mainz / Universitätsmedizin	de
dc.language.iso	eng	de
dc.rights	CC BY	*
dc.rights.uri	https://creativecommons.org/licenses/by/4.0/	*
dc.subject.ddc	500 Naturwissenschaften	de_DE
dc.subject.ddc	500 Natural sciences and mathematics	en_GB
dc.title	Absolute and relative quantification of RNA modifications via biosynthetic isotopomers	en_GB
dc.type	Zeitschriftenaufsatz	de
dc.identifier.doi	http://doi.org/10.25358/openscience-8002	-
jgu.type.dinitype	article	en_GB
jgu.type.version	Published version	de
jgu.type.resource	Text	de
jgu.organisation.department	FB 09 Chemie, Pharmazie u. Geowissensch.	de
jgu.organisation.number	7950	-
jgu.organisation.name	Johannes Gutenberg-Universität Mainz	-
jgu.rights.accessrights	openAccess	-
jgu.journal.title	Nucleic acids research	de
jgu.journal.volume	42	de
jgu.journal.issue	18	de
jgu.pages.alternative	e142	de
jgu.publisher.year	2014	-
jgu.publisher.name	Oxford Univ. Press	de
jgu.publisher.place	Oxford	de
jgu.publisher.uri	http://dx.doi.org/10.1093/nar/gku733	de
jgu.publisher.issn	1362-4962	de
jgu.publisher.issn	0305-1048	de
jgu.organisation.place	Mainz	-
jgu.subject.ddccode	500	de
opus.date.modified	2018-08-08T08:02:05Z
opus.subject.dfgcode	00-000
opus.organisation.string	FB 09: Chemie, Pharmazie und Geowissenschaften: Institut für Pharmazie	de_DE
opus.identifier.opusid	26880
opus.institute.number	0908
opus.metadataonly	false
opus.type.contenttype	Keine	de_DE
opus.type.contenttype	None	en_EN
opus.affiliated	Schneider, Dirk
opus.affiliated	Helm, Mark
jgu.publisher.doi	10.1093/nar/gku733	de
jgu.organisation.ror	https://ror.org/023b0x485	-
Appears in collections:	DFG-OA-Publizieren (2012 - 2017)

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