Journal
/
Génétique
/
Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
Journal JoVE
Génétique
Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Journal JoVE Génétique
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

•

10:23 min

•

January 19, 2017

•

, , , ,
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

Chapitres

  • 00:05Titre
  • 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

Traduction automatique

Traduction automatique

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

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

Perturbations of Circulating miRNAs in Irritable Bowel Syndrome Detected Using a Multiplexed High-throughput Gene Expression Platform

Detection of Copy Number Alterations Using Single Cell Sequencing

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols

Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish

Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput

An Optogenetic Method to Control and Analyze Gene Expression Patterns in Cell-to-cell Interactions

Using Mouse Oocytes to Assess Human Gene Function During Meiosis I

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Exploring the Root Microbiome: Extracting Bacterial Community Data from the Soil, Rhizosphere, and Root Endosphere

Read Article