title: Comprehensive qPCR profiling of gene expression in single neuronal cells.
abstract: A major challenge in neuronal stem cell biology lies in characterization of lineage-specific reprogrammed human neuronal cells, a process that necessitates the use of an assay sensitive to the single-cell level. Single-cell gene profiling can provide definitive evidence regarding the conversion of one cell type into another at a high level of resolution. The protocol we describe uses Fluidigm Biomark dynamic arrays for high-throughput expression profiling from single neuronal cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment, which can be completed within 2-3 d. The protocol enables simple and cost-effective profiling of several hundred transcripts from a single cell, and it could have numerous utilities.
authors.affiliation: Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, California, USA. citri@stanford.edu
authors.forename: Ami
authors.initials: A
authors.lastname: Citri
authors.forename: Zhiping P
authors.initials: ZP
authors.lastname: Pang
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Südhof
authors.forename: Marius
authors.initials: M
authors.lastname: Wernig
authors.forename: Robert C
authors.initials: RC
authors.lastname: Malenka
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D044085:Microfluidics; D009474:Neurons / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D059010:Single-Cell Analysis
chemical_list: D012333:RNA, Messenger