Isoform discovery by targeted cloning, 'deep-well' pooling and parallel sequencing

Kourosh Salehi-Ashtiani, Xinping Yang, Adnan Derti, Weidong Tian, Tong Hao, Chenwei Lin, Kathryn Makowski, Lei Shen, Ryan R. Murray, David Szeto, Nadeem Tusneem, Douglas R. Smith, Michael E. Cusick, David E. Hill, Frederick P. Roth, Marc Vidal

Research output: Contribution to journalArticlepeer-review

Abstract

Describing the 'ORFeome' of an organism, including all major isoforms, is essential for a system-level understanding of any species; however, conventional cloning and sequencing approaches are prohibitively costly and labor-intensive. We describe a potentially genome-wide methodology for efficiently capturing new coding isoforms using reverse transcriptase (RT)-PCR recombinational cloning, 'deep-well' pooling and a next-generation sequencing platform. This ORFeome discovery pipeline will be applicable to any eukaryotic species with a sequenced genome.

Original languageEnglish (US)
Pages (from-to)597-600
Number of pages4
JournalNature methods
Volume5
Issue number7
DOIs
StatePublished - Jul 2008

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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