Abstract
Single-cell RNA-seq technology allows for the identification of heterogeneous cell populations, measures stochastic gene expressions, and identifies highly variable genes. Thus, with this technology it is possible to identify relevant pathways involved in development or in disease progression. Herein, we describe a protocol to capture and process single-cell transcriptomes that will be used for RNA sequencing. This chapter discusses the use of the Fluidigm C1 System and Integrated Fluidic Circuit microfluidics system, TapeStation 4200, SMART-Seq v4, Nextera XT Library Preparation Kit, and AMPure XP beads.
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References
Behjati S, Tarpey PS (2013) What is next generation sequencing? Arch Dis Child Educ Pract Ed 98:236–238
Bengtsson M, Ståhlberg A, Rorsman P, Kubista M (2005) Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levels. Genome Res 15:1388–1392
Islam S et al (2011) Characterization of the single-cell transcriptional landscape by highly multiplex RNA-seq. Genome Res 21:1160–1167
Gross A et al (2015) Technologies for single-cell isolation. Int J Mol Sci 16:16897–16919
Welzel G, Seitz D, Schuster S (2015) Magnetic-activated cell sorting (MACS) can be used as a large-scale method for establishing zebrafish neuronal cell cultures. Sci Rep 5:7959
Datta S et al (2015) Laser capture microdissection: big data from small samples. Histol Histopathol 30:1255–1269
Bhagat AAS et al (2010) Microfluidics for cell separation. Med Biol Eng Comput 48:999–1014
Acknowledgment
The authors thank members of the Daadi laboratory for the helpful support and suggestions. This work was supported by the Worth Family Fund, the Perry & Ruby Stevens Charitable Foundation and the Robert J., Jr. and Helen C. Kleberg Foundation, the NIH primate center base grant (Office of Research Infrastructure Programs/OD P51 OD011133), and the National Center for Advancing Translational Sciences, National Institutes of Health, through Grant UL1 TR001120.
Disclosures: Dr. Marcel M. Daadi is founder of the biotech company NeoNeuron.
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Kim, J., Daadi, M.M. (2019). Single-Cell Library Preparation of iPSC-Derived Neural Stem Cells. In: Daadi, M. (eds) Neural Stem Cells. Methods in Molecular Biology, vol 1919. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9007-8_10
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DOI: https://doi.org/10.1007/978-1-4939-9007-8_10
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9005-4
Online ISBN: 978-1-4939-9007-8
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