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. 2019 Jan 5;10(1):26. doi: 10.3390/genes10010026

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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To produce stable isotope-labeled internal standards (SILIS), Escherichia coli (E. coli) or Saccharomyces cerevisiae (S. cerevisiae) were cultured in isotope labeled media using nutrients with the highest isotope-purity possible. The total RNA was isolated and then separated into transfer RNA (tRNA) or large/ribosomal RNA through size exclusion chromatography. (* indicates 5.8 and 5S rRNA and # phenol) The different RNAs were enzymatically digested into nucleosides and analyzed by liquid chromatography–mass spectrometry (LC–MS/MS).