Quantitative end-point RT-PCR gene expression measurement using the Agilent 2100 Bioanalyzer and standardized RT-PCR (5988-3674EN)
نویسنده
چکیده
This application note describes how the Agilent 2100 bioanalyzer can be used in combination with Standardized RT (StaRT)-PCR reagents from Gene Express National Enterprises (G.E.N.E.), Inc. (Huntsville, AL, USA) to obtain highly reproducible, standardized gene expression measurement with RT-PCR. The internal standards used in StaRT-PCR control for variable loading of PCR products into electrophoresis channel, and allow quantitative gene expression measurement at the endpoint of PCR, without real-time analysis. Gene expression measurement of 15 genes was more reproducible with the Agilent 2100 bioanalyzer than with the ABI Prism 310 Genetic Analyzer or agarose gel electrophoresis methods. The mean coefficient of variance (CV) for these three methods was 0.26, 0.39 and 0.50, respectively. Erin L. Crawford, M.S. Kristy A. Warner, Ph.D. David A. Weaver, Ph.D. James C. Willey, M.D.
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