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Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
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JoVE Revista Genética
Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

DOI:
10.3791/54858-v

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10:23 min

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January 19, 2017

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, , , ,
1Unit for Lymphopoieseis, Immunology Department, INSERM U1223, University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur,Institut Pasteur, 2Center for Translational Science,Institut Pasteur, INSERM UMS20

Capítulos

  • 00:05Título
  • 00:40Preparing a 96-well Single-cell Sorting Plate
  • 01:55Single-cell Fluorescence-activated Cell Sorting (FACS)
  • 03:02Pre-amplification
  • 03:38Preparing a 96-well Sample Plate
  • 04:23Preparing a 96-well Assay Plate
  • 04:51Preparing and Running the Single-cell Gene-expression Chip
  • 06:52Data Analysis
  • 08:04Results: Single-cell Multiplex RT-qPCR Identifies Cell Heterogeneity of the Liver ILC Compartment
  • 09:33Conclusion

Summary

Traducción Automática

Traducción Automática

This protocol describes how to assess the expression of a large array of genes at the clonal level. Single-cell RT-qPCR produces highly reliable results with a strong sensitivity for hundreds of samples and genes.

Tags

Single-cell Gene ExpressionMultiplex RT-qPCRHeterogeneityRare Lymphoid PopulationsPre-amplificationAssay MixFACSInnate Lymphoid CellsNo-input ControlCDNA DilutionReverse TranscriptionPre-amplification

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