Single-Cell Library Preparation of iPSC-Derived Neural Stem Cells

Jeffrey Kim; Marcel M Daadi

doi:10.1007/978-1-4939-9007-8_10

Single-Cell Library Preparation of iPSC-Derived Neural Stem Cells

Methods Mol Biol. 2019:1919:129-143. doi: 10.1007/978-1-4939-9007-8_10.

Authors

Jeffrey Kim¹, Marcel M Daadi^{2

3}

Affiliations

¹ Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX, USA.
² Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX, USA. mdaadi@txbiomed.org.
³ Department of Radiology, Research Imaging Institute, Cell Systems and Anatomy, Long School of Medicine, University of Texas Health Science Center at San Antonio , San Antonio, TX, USA. mdaadi@txbiomed.org.

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.

Keywords: C1; Fluidigm; Library preparation; Neural stem cells; Next-generation sequencing; Nextera XT; RNA-seq; SMART-Seq; Single cell; Single-cell RNA-seq.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Gene Library*
High-Throughput Nucleotide Sequencing
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism*
Neural Stem Cells / cytology*
Neural Stem Cells / metabolism*
Single-Cell Analysis*
Transcriptome
Workflow

Abstract

Publication types

MeSH terms

Grants and funding