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Keywords {🔍}

celseq, cells, cell, file, fig, genes, sequencing, additional, singlecell, article, number, methods, protocol, expression, method, analysis, data, sensitivity, mouse, gfp, smartseq, rna, primer, library, transcripts, original, detected, step, efficiency, reads, performed, single, cdna, umi, primers, pubmed, rnaseq, gene, figure, pipeline, sample, comparison, ligation, illumina, synthesis, read, barcodes, fluidigm, variation, google,

Topics {✒️}

dmem/10 % fbs/1 % pen-strep/1 % l-glutamine single-cell rna-seq analysis high-throughput sequencing data enabling highly-multiplexed analyses orit rozenblatt-rosen enabling highly-multiplexed analysis single-cell transcriptomics applied itai yanai highly multiplex rna-seq medical research israel-canada fluidigm c1-enabled cell-barcoding truseq small-rna kit single-cell rna-seq single-cell transcriptional landscape double stranded dna single-cell sequencing experiments mouse fibroblast cells mouse dendritic cells fast gapped-read alignment single-cell transcriptomics template-switch step full-length mrna-seq cyclinb1-gfp fusion reporter produces full-length transcripts avoid low-complexity barcodes /yanailab/cel-seq-pipeline multiplexed linear amplification reverse transcription cel-seq2 cel-seq article download pdf messageamp ii enzyme recessive fibroblast variants illumina adaptor directly library preparation step original cel-seq protocol hadassah medical center single sequencing error single-cell levels detected ercc spike-ins detailed cel-seq2 protocol single-cell method privacy choices/manage cookies ligation step technion genome center full transcript sequencing showed high coefficient superscript ii single library construction template switching mechanism reverse-transcribed mrna

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         description:Single-cell transcriptomics requires a method that is sensitive, accurate, and reproducible. Here, we present CEL-Seq2, a modified version of our CEL-Seq method, with threefold higher sensitivity, lower costs, and less hands-on time. We implemented CEL-Seq2 on Fluidigm’s C1 system, providing its first single-cell, on-chip barcoding method, and we detected gene expression changes accompanying the progression through the cell cycle in mouse fibroblast cells. We also compare with Smart-Seq to demonstrate CEL-Seq2’s increased sensitivity relative to other available methods. Collectively, the improvements make CEL-Seq2 uniquely suited to single-cell RNA-Seq analysis in terms of economics, resolution, and ease of use.
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               type:PostalAddress
            type:Organization
      name:Itai Yanai
      affiliation:
            name:Technion – Israel Institute of Technology
            address:
               name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
      name:Department of Developmental Biology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem, Israel
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel
      name:Klarman Cell Observatory, Broad Institute of Harvard and MIT, Cambridge, USA
      name:Department of Biology, MIT, Cambridge, USA
      name:Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, USA
      name:Fluidigm Corporation, South San Francisco, USA
      name:Fluidigm Corporation, South San Francisco, USA
      name:Klarman Cell Observatory, Broad Institute of Harvard and MIT, Cambridge, USA
      name:Department of Biology, MIT, Cambridge, USA
      name:Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, USA
      name:Department of Developmental Biology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem, Israel
      name:Klarman Cell Observatory, Broad Institute of Harvard and MIT, Cambridge, USA
      name:Department of Biology, MIT, Cambridge, USA
      name:Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, USA
      name:Department of Biology, Technion – Israel Institute of Technology, Haifa, Israel

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