title: Transcriptome Analysis at the Single-Cell Level Using SMART Technology.
abstract: RNA sequencing (RNA-seq) is a powerful method for analyzing cell state, with minimal bias, and has broad applications within the biological sciences. However, transcriptome analysis of seemingly homogenous cell populations may in fact overlook significant heterogeneity that can be uncovered at the single-cell level. The ultra-low amount of RNA contained in a single cell requires extraordinarily sensitive and reproducible transcriptome analysis methods. As next-generation sequencing (NGS) technologies mature, transcriptome profiling by RNA-seq is increasingly being used to decipher the molecular signature of individual cells. This unit describes an ultra-sensitive and reproducible protocol to generate cDNA and sequencing libraries directly from single cells or RNA inputs ranging from 10 pg to 10 ng. Important considerations for working with minute RNA inputs are given. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Takara Bio USA, Inc. (formerly Clontech Laboratories, Inc.), Mountain View, California.
authors.forename: Rachel N
authors.initials: RN
authors.lastname: Fish
authors.affiliation: Takara Bio USA, Inc. (formerly Clontech Laboratories, Inc.), Mountain View, California.
authors.forename: Magnolia
authors.initials: M
authors.lastname: Bostick
authors.affiliation: Takara Bio USA, Inc. (formerly Clontech Laboratories, Inc.), Mountain View, California.|Current affiliation: 23andMe, Mountain View, California.
authors.forename: Alisa
authors.initials: A
authors.lastname: Lehman
authors.affiliation: Takara Bio USA, Inc. (formerly Clontech Laboratories, Inc.), Mountain View, California.
authors.forename: Andrew
authors.initials: A
authors.lastname: Farmer
mesh_terms: D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods*; D059467:Transcriptome
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D012313:RNA
keywords: RNA-seq; gene expression; next-generation sequencing; single cell; transcriptome