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Bioengineering

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JoVE Journal Bioengineering
High Throughput MicroRNA Profiling
 

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Article DOI: 10.3791/2552-v • 07:27 min • August 3rd, 2011
August 3rd, 2011 •
1,2,3, 1,2,4, 5, 1,2,6, 1,2,3,7
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

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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 Profiling Multiplex QRT-PCR Fluidigm Dynamic ArrayTM IFCs MiRNAs Quantification MicroRNA Levels Wild-type Small RNA Deficient Mouse Embryonic Stem Cells (mESC) Accuracy Robustness Optimized Method Purifying Primers Singleplex Real-time Detection Microfluidic Chip Nanoliter Volumes Reagent Costs High Throughput MiRNA Expression Profiling
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