Amplification-free digital gene expression profiling from minute cell quantities.
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.
Ozsolak
Fatih
F
Helicos BioSciences Corporation, Cambridge, Massachusetts, USA. fatihozsolak@gmail.com
Ting
David T
DT
Wittner
Ben S
BS
Brannigan
Brian W
BW
Paul
Suchismita
S
Bardeesy
Nabeel
N
Ramaswamy
Sridhar
S
Milos
Patrice M
PM
Haber
Daniel A
DA
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
D012333:RNA, Messenger; D011071:Poly T