期刊
NATURE METHODS
卷 5, 期 7, 页码 597-600出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.1224
关键词
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资金
- NHGRI NIH HHS [HG003224, R01 HG003224-03, R01 HG003224-01A2, R01 HG003224, R01 HG003224-02] Funding Source: Medline
- NHLBI NIH HHS [HL081341, U01 HL081341-01, U01 HL081341-02, U01 HL081341-03, U01 HL081341] Funding Source: Medline
- NIDDK NIH HHS [DK070078, T90 DK070078-03, T90 DK070078] Funding Source: Medline
- NINDS NIH HHS [R01 NS054052, R01 NS054052-02, NS054052, R01 NS054052-01A1] Funding Source: Medline
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.
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