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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.

SUBMITTER: Ozsolak F

PROVIDER: S-EPMC2975601 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature

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Amplification-free digital gene expression profiling from minute cell quantities.

Ozsolak Fatih F 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

Nature methods 20100718 8

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 quant ...[more]

PMID: 20639869

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