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CEL-Seq2-Single-Cell RNA Sequencing by Multiplexed Linear Amplification.

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  • 1Institute for Computational Medicine, NYU School of Medicine, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2019
PubMed
Summary
This summary is machine-generated.

We describe CEL-Seq2, a cost-effective and sensitive protocol for single-cell RNA sequencing. This method uses linear amplification and early barcoding for analyzing cell populations without special equipment.

Keywords:
3′ sequencingGene expressionIn vitro transcriptionSingle-cell RNA amplificationSingle-cell transcriptomics

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Single-cell RNA sequencing (scRNA-seq) offers unprecedented resolution into cellular heterogeneity.
  • CEL-Seq was a pioneering method utilizing linear RNA amplification for scRNA-seq.
  • Existing methods often require specialized equipment or complex procedures.

Purpose of the Study:

  • To present an optimized protocol for CEL-Seq2, a refined version of the original CEL-Seq method.
  • To demonstrate the feasibility of CEL-Seq2 for analyzing sorted cells.
  • To provide a cost-effective and accessible scRNA-seq protocol.

Main Methods:

  • The protocol involves linear RNA amplification and early barcoding.
  • It is designed for use with sorted cells.
  • The procedure is streamlined to minimize the need for specialized laboratory equipment.

Main Results:

  • CEL-Seq2 maintains the sensitivity and cost-effectiveness of the original CEL-Seq.
  • The protocol is robust and easy to perform on sorted cell populations.
  • Successful application of CEL-Seq2 enables detailed analysis of cellular gene expression.

Conclusions:

  • CEL-Seq2 provides a valuable and accessible tool for single-cell RNA sequencing.
  • The method facilitates the study of cell populations without requiring specialized infrastructure.
  • This protocol advances the application of scRNA-seq in various research fields.