The Use of the Fluidigm C1 for RNA Expression Analyses of Single Cells

Current Protocols in Molecular Biology
Daniel M DeLaughter

Abstract

Understanding the transcriptional heterogeneity that occurs on the level of a single cell is critical to understanding the gene-regulatory mechanisms underlying development and disease. Population-level whole-transcriptome profiling approaches average gene expression across thousands to millions of cells and are unable to delineate the transcriptional signature of individual cells. Considerable biological heterogeneity between individual cells arises from differences in cell lineage, environment, or response to stimulus. The development of single-cell RNA sequencing (RNA-seq) enabled a high-resolution and unbiased analysis of cell transcriptomes. This unit describes a procedure utilizing an automated microfluidic platform, the Fluidigm C1 system, to simultaneously isolate dozens of single cells in a size- and shape-dependent manner. The microfluidic platform processes cells in individual nanoliter-scale reactions to convert their contents into double-stranded cDNA. This cDNA is used to make dual-indexed libraries using the Illumina Nextera XT library preparation kit for eventual RNA-seq analysis. © 2018 by John Wiley & Sons, Inc.

References

Apr 8, 2009·Nature Methods·Fuchou TangM Azim Surani
Sep 24, 2013·Nature Methods·Simone PicelliRickard Sandberg
Dec 24, 2013·Nature Methods·Saiful IslamSten Linnarsson
May 23, 2015·Molecular Cell·Aleksandra A KolodziejczykSarah A Teichmann
Jan 17, 2017·Nature Communications·Grace X Y ZhengJason H Bielas
Feb 19, 2017·Molecular Cell·Christoph ZiegenhainWolfgang Enard
Jun 8, 2017·Frontiers in Genetics·Alessandra Dal MolinBarbara Di Camillo

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell-Type Specific Viral Vectors

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.

Cell-Type-Specific Viral Vectors (ASM)

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.

Cell-Type-Specific Viral Vectors

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.

Related Papers

Current Protocols in Molecular Biology
Rachel N FishAndrew Farmer
Current Protocols in Molecular Biology
John J TrombettaAviv Regev
Current Protocols in Mouse Biology
Wanze ChenBart Deplancke
Current Protocols in Molecular Biology
Nan Fang, Rumeysa Akinci-Tolun
Environmental Science & Technology
© 2022 Meta ULC. All rights reserved