title: Dextran sodium sulfate inhibition of real-time polymerase chain reaction amplification: a poly-A purification solution.
abstract: BACKGROUND
  Dextran sulfate sodium (DSS) induces experimental colitis and promotes colitis-associated cancer in rodents. Here we document potent inhibition of real-time quantitative polymerase chain reaction (qPCR) using cDNA from DSS-exposed mouse tissues, which complicates gene expression analysis.
  
  
  METHODS
  We characterize DSS inhibition of qPCR in-vitro and in a wide array of murine tissues following ingestion of DSS. We examine different approaches to RNA purification prior to cDNA synthesis in order to optimize real-time polymerase chain reaction amplification and gene expression analysis.
  
  
  RESULTS
  DSS inhibits qPCR amplification of cDNA between 1 and 10 nM. Orally administered DSS interferes with qPCR amplification of cDNA derived from multiple tissues. Poly-A purification of DSS-exposed RNA allows reliable and cost-effective gene expression analysis in DSS-exposed tissue.
  
  
  CONCLUSIONS
  DSS is a potent inhibitor of real-time qPCR amplification and interferes with tissue-specific gene expression analysis in DSS-exposed mice. Poly-A purification of tissue-derived RNA results in reliable and cost-effective gene expression analysis in DSS-exposed mice.
authors: Gastroenterology Division, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors: T A
authors: TA
authors: Kerr
authors: M A
authors: MA
authors: Ciorba
authors: H
authors: H
authors: Matsumoto
authors: V R T
authors: VR
authors: Davis
authors: J
authors: J
authors: Luo
authors: S
authors: S
authors: Kennedy
authors: Y
authors: Y
authors: Xie
authors: A
authors: A
authors: Shaker
authors: B K
authors: BK
authors: Dieckgraefe
authors: N O
authors: NO
authors: Davidson
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