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Related Concept Videos

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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Related Experiment Video

Updated: Sep 6, 2025

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
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Single-Cell RNA Sequencing Analysis Using Fluidigm C1 Platform for Characterization of Heterogeneous Transcriptomes.

Jinhong Kim1, Paola A Marignani2,3

  • 1Department of Biochemistry and Molecular Biology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|June 23, 2022
PubMed
Summary

Single-cell RNA sequencing (scRNA-seq) offers high-resolution transcriptome analysis. This study details a workflow for 3'-end enriched cDNA libraries using the Fluidigm C1 system and validates differential gene expression.

Keywords:
Differential gene expressionFluidigm C1 systemsProtein expression analysisValidationqRT-PCRscRNA-seq

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Single-cell RNA sequencing (scRNA-seq) enables the study of cellular heterogeneity.
  • Automated platforms like the Fluidigm C1 streamline single-cell sample preparation.
  • Accurate gene expression analysis is crucial for understanding cellular function.

Purpose of the Study:

  • To present a detailed workflow for scRNA-seq using 3 e2 -end enriched cDNA libraries.
  • To introduce methods for validating differential gene expression identified through scRNA-seq.
  • To optimize the use of the Fluidigm C1 system for scRNA-seq applications.

Main Methods:

  • Utilized the Fluidigm C1 system for automated single-cell capture, lysis, and cDNA synthesis.
  • Prepared 3 e2 -end enriched cDNA libraries suitable for next-generation sequencing (NGS).
  • Implemented strategies for the statistical validation of differentially expressed genes.

Main Results:

  • Established a robust and reproducible protocol for scRNA-seq library preparation.
  • Demonstrated the effectiveness of the Fluidigm C1 system for generating high-quality scRNA-seq data.
  • Validated differential gene expression findings through established bioinformatics approaches.

Conclusions:

  • The presented workflow facilitates efficient and reliable scRNA-seq analysis.
  • The Fluidigm C1 system is a valuable tool for single-cell transcriptomics.
  • The validation strategies ensure the accuracy of differential gene expression findings in scRNA-seq studies.