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Cell Rep. 2012 Sep 27;2(3):666-73. doi: 10.1016/j.celrep.2012.08.003. Epub 2012 Aug 30.

CEL-Seq: single-cell RNA-Seq by multiplexed linear amplification.

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  • 1Department of Biology, Technion-Israel Institute of Technology, Haifa 32000, Israel.

Abstract

High-throughput sequencing has allowed for unprecedented detail in gene expression analyses, yet its efficient application to single cells is challenged by the small starting amounts of RNA. We have developed CEL-Seq, a method for overcoming this limitation by barcoding and pooling samples before linearly amplifying mRNA with the use of one round of in vitro transcription. We show that CEL-Seq gives more reproducible, linear, and sensitive results than a PCR-based amplification method. We demonstrate the power of this method by studying early C. elegans embryonic development at single-cell resolution. Differential distribution of transcripts between sister cells is seen as early as the two-cell stage embryo, and zygotic expression in the somatic cell lineages is enriched for transcription factors. The robust transcriptome quantifications enabled by CEL-Seq will be useful for transcriptomic analyses of complex tissues containing populations of diverse cell types.

Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.

PMID:
22939981
[PubMed - indexed for MEDLINE]
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