Single-Cell Library Preparation of iPSC-Derived Neural Stem Cells
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.
Key wordsSingle cell Neural stem cells Fluidigm C1 Library preparation RNA-seq Single-cell RNA-seq Nextera XT Next-generation sequencing SMART-Seq
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.