title: Highly consistent, fully representative mRNA-Seq libraries from ten nanograms of total RNA.
abstract: Preparation of an Illumina sequencing library for gene expression analysis (mRNA-Seq) requires microgram amounts of starting total RNA or PCR-based amplification. Here we describe a protocol based on T7 linear RNA amplification that does not introduce significant bias, requires only 10 ng total RNA, and generates a directional, fully representative, whole-transcript mRNA-Seq Illumina library that is highly consistent across over three orders of magnitude of input RNA.
authors.affiliation: Morgridge Institute for Research, Madison, WI 53706, USA.
authors.forename: Srikumar
authors.initials: S
authors.lastname: Sengupta
authors.forename: Victor
authors.initials: V
authors.lastname: Ruotti
authors.forename: Jennifer
authors.initials: J
authors.lastname: Bolin
authors.forename: Angela
authors.initials: A
authors.lastname: Elwell
authors.forename: Alvaro
authors.initials: A
authors.lastname: Hernandez
authors.forename: James
authors.initials: J
authors.lastname: Thomson
authors.forename: Ron
authors.initials: R
authors.lastname: Stewart
mesh_terms: D002460:Cell Line; D053595:Embryonic Stem Cells / Q000502:physiology; D020869:Gene Expression Profiling; D015723:Gene Library; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D017423:Sequence Analysis, RNA
chemical_list: D012333:RNA, Messenger; D012313:RNA