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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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August 03, 2011
DOI:
1The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research,University of California San Francisco , 2Center for Reproductive Sciences,University of California San Francisco , 3Department of Urology,University of California San Francisco , 4Department of Cell and Tissue Biology,University of California San Francisco , 5Fluidigm Corporation,Fluidigm Corporation , 6Department of Obstetrics and Gynecology,Hadassah-Hebrew University Medical Center, 7UCSF – Helen Diller Family Comprehensive Cancer Center,University of California San Francisco
Summary
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Here we describe an optimized multiplex reverse transcriptase quantitative PCR (qRT-PCR) protocol in combination with a microfluidic platform as a cost and time effective high-throughput screening tool for microRNA (miRNA) expression levels, especially when working with limited amounts of sample.
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High Throughput MicroRNA ProfilingMultiplex QRT-PCRFluidigm Dynamic ArrayTM IFCsMiRNAsQuantificationMicroRNA LevelsWild-typeSmall RNA Deficient Mouse Embryonic Stem Cells (mESC)AccuracyRobustnessOptimized MethodPurifying PrimersSingleplex Real-time DetectionMicrofluidic ChipNanoliter VolumesReagent CostsHigh Throughput MiRNA Expression Profiling
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Moltzahn, F., Hunkapiller, N., Mir, A. A., Imbar, T., Blelloch, R. High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs. J. Vis. Exp. (54), e2552, doi:10.3791/2552 (2011).
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