title: Single-cell PCR profiling of gene expression in hematopoiesis.
abstract: Single-cell analysis of gene expression offers the possibility of exploring cellular and molecular heterogeneity in stem and developmental cell systems, including cancer, to infer routes of cellular specification and their respective gene regulatory modules. PCR-based technologies, although limited to the analysis of a predefined set of genes, afford a cost-effective balance of throughput and biological information and have become a method of choice in stem cell laboratories. Here we describe an experimental and analytical protocol based on the Fluidigm microfluidics platform for the simultaneous expression analysis of 48 or 96 genes in multiples of 48 or 96 cells. We detail wet laboratory procedures and describe clustering, principal component analysis, correlation, and classification tools for the inference of cellular pathways and gene networks.
authors: Stem Cell Laboratory, University College London Cancer Institute, 72 Huntley Street, WC1E 6BT, London, UK.
authors: José
authors: J
authors: Teles
authors: Tariq
authors: T
authors: Enver
authors: Cristina
authors: C
authors: Pina
mesh_terms: D000818:Animals; D016923:Cell Death; D002460:Cell Line; D002469:Cell Separation; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D006410:Hematopoiesis / Q000235:genetics*; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D025341:Principal Component Analysis; D011786:Quality Control; D048348:Reverse Transcription; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D017931:DNA Primers