Rapid removal of dextran sulfate sodium from tissue RNA preparations for measurement of inflammation biomarkers

Anthony F. Juritsch, Régis Moreau

Research output: Contribution to journalArticle

Abstract

Dextran sulfate sodium (DSS) present in the tissues of DSS-treated laboratory animals inhibits quantitative real-time qPCR (RT-qPCR) and thus may be source of experimental errors. A recent systematic review concluded that the reporting of experimental method was insufficient in a majority of DSS studies and contributed to the poor reproducibility of experiments. Here we compared two DSS cleanup protocols applied to mouse tissue RNA preparations based on silica membrane spin column and lithium chloride precipitation. In absence of cleanup, exogenous DSS significantly inhibited reverse transcription and cDNA amplification at concentrations of 5 × 10−3 g/L and above during the quantification of IL8 mRNA levels in THP-1 macrophages. Silica membrane spin columns removed DSS from mouse RNA preparations and eliminated DSS-induced inhibition of qPCR. Mouse RNA isolated from DSS-treated tissues and purified with silica membrane spin columns was suitable for RT-qPCR and assessment of inflammatory biomarkers.

Original languageEnglish (US)
Pages (from-to)18-24
Number of pages7
JournalAnalytical Biochemistry
Volume579
DOIs
StatePublished - Aug 15 2019

Keywords

  • Colitis
  • Cytokine
  • Gut
  • Inflammatory bowel disease
  • mRNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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