title: Amplification-free digital gene expression profiling from minute cell quantities.
abstract: Generating reliable expression profiles from minute cell quantities is critical for scientific discovery and potential clinical applications. Here we present low-quantity digital gene expression (LQ-DGE), an amplification-free approach involving capture of poly(A)(+) RNAs from cellular lysates onto poly(dT)-coated sequencing surfaces, followed by on-surface reverse transcription and sequencing. We applied LQ-DGE to profile malignant and nonmalignant mouse and human cells, demonstrating its quantitative power and potential applicability to archival specimens.
authors.affiliation: Helicos BioSciences Corporation, Cambridge, Massachusetts, USA. fatihozsolak@gmail.com
authors.forename: Fatih
authors.initials: F
authors.lastname: Ozsolak
authors.forename: David T
authors.initials: DT
authors.lastname: Ting
authors.forename: Ben S
authors.initials: BS
authors.lastname: Wittner
authors.forename: Brian W
authors.initials: BW
authors.lastname: Brannigan
authors.forename: Suchismita
authors.initials: S
authors.lastname: Paul
authors.forename: Nabeel
authors.initials: N
authors.lastname: Bardeesy
authors.forename: Sridhar
authors.initials: S
authors.lastname: Ramaswamy
authors.forename: Patrice M
authors.initials: PM
authors.lastname: Milos
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Haber
mesh_terms: D000818:Animals; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D051379:Mice; D046228:Microarray Analysis / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011071:Poly T / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D048348:Reverse Transcription; D017422:Sequence Analysis, DNA
chemical_list: D012333:RNA, Messenger; D011071:Poly T