Dextran sodium sulfate inhibition of real-time polymerase chain reaction amplification: a poly-A purification solution.

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.

Demonstration of room-temperature laser action at 2.5 mum from Cr(2+):Cd(0.85)Mn(0.15)Te.

Room-temperature laser action from Cr(2+)-doped Cd(0.85)Mn(0.15)Te has been demonstrated for what is believed to be the first time. We achieved pulsed laser operation centered at ~2.5mu m by pumping into the mid-infrared absorption band of Cr(2+) ions by use of the 1.907- mum output of a H(2) Raman-shifted Nd:YAG laser. The output of the free-running Cr(2+):Cd(0.85)Mn(0.15)Te laser had a width of ~50 nm (FWHM), and the slope efficiency was calculated to be 5.5% under nonoptimum conditions.