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miniaturization and automation of CEL-Seq2 and SMART-Seq2 using the mosquito liquid handler
Poster Title: miniaturization and automation of CEL-Seq2 and SMART-Seq2 using the mosquito liquid handler
Submitted on 10 Jan 2020
Author(s): Josip Herman1, Jon Penterman2, Sagar1, Andreas Diefenbach3, Antigoni Triantafyllopoulou4, Anne F. Hammerstein5, Joby Jenkins5, Dominic Grün1, Stuart S. Levine2 and Klaus Hentrich5
Affiliations: 1 Max Planck Institute of Immunobiology and Epigenetics, 79108 Freiburg, Germany; 2 BioMicro Center, Massachussets Institute of Technology, Cambridge, MA 02139, USA; 3 Institute of Medical Microbiology and Hygiene, University of Mainz Medical Center, 55131 Mainz, Germany; 4 Department of Rheumatology and Clinical Immunology, Medical Center–University of Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany; 5 SPT Labtech, Melbourn Science Park, Melbourn, Hertfordshire SG8 6HB, UK
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Poster Information
Abstract: Plate-based workflows with flow-sorted single cells allow the analysis of rare cell types and subpopulations characterised by cell surface markers. Automation and miniaturisation are key to make such single cell sequencing studies practical and cost effective at high throughput. Liquid handling steps on robotic platforms must be accurate and robust to avoid any loss of material, prevent cross-contamination, and limit the introduction of technical

Here we present the use of SPT Labtech’s mosquito® liquid handlers to automate and miniaturise two different single cell RNA-seq protocols with FACS-sorted cells in 384-well plates.
Summary: automation and miniaturisation of CEL-Seq2 cDNA
synthesis with mosquito LV increased throughput,
improved process consistency and reduced cost
~5-fold, while maintaining high sensitivity and
MIT BioMicro Center’s SMART-Seq2 workflow with
mosquito HV overcomes challenges with cost,
throughput and diversity of samples, and delivers
the required data quality for high-throughput single-cell
References: 1.Herrtwich et al., 2016. DNA Damage Signaling Instructs Polyploid Macrophage Fate in Granulomas. Cell 167(5), 1264-1280.
2.Herman et al., 2017. FateID infers cell fate bias in multipotent progenitors from single-cell RNA-seq data. submitted / bioRxiv preprint:
3.Hashimshony et al., 2016. CEL-Seq2: sensitive highly-multiplexed single-cell RNASeq. Genome Biology 17:77.
4.Picelli et al., 2014. Full-length RNA-seq from single cells using Smart-Seq2. Nature Protocols 9(1):171-81.
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