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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
Chapters
- 00:05Title
- 00:55RNA Concentration
- 02:03Gel Purification of pre-PCR Product
- 03:46Preparing the 96.96 Dynamic Array IFC, the Samples, and the 13x Assays
- 05:25qRT-PCR Analysis of miRNA Expression in Mouse ESC and Human Serum
- 06:10qRT-PCR Analysis
- 07:04Conclusion
A continuación se describe un reverso transcriptasa optimizado multiplex PCR cuantitativa (QRT-PCR) en combinación con el protocolo de una plataforma de microfluidos como un costo y el tiempo efectivo instrumento de cribado de alto rendimiento para los microRNA (miRNA) los niveles de expresión, especialmente cuando se trabaja con cantidades limitadas de muestras.
Tags
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
