Open Access Method

A genome annotation-driven approach to cloning the human ORFeome

John E Collins1, Charmain L Wright1, Carol A Edwards13, Matthew P Davis1, James A Grinham1, Charlotte G Cole1, Melanie E Goward1, Begoña Aguado24, Meera Mallya25, Younes Mokrab16, Elizabeth J Huckle1, David M Beare1 and Ian Dunham1*

Author affiliations

1 The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, UK

2 MRC Rosalind Franklin Centre for Genomics Research (formerly MRC UK Human Genome Mapping Resource Centre), Hinxton, Cambridge, CB10 1SB, UK

3 Current address: Department of Anatomy, University of Cambridge, Downing Street, Cambridge, CB2 3DY, UK

4 Current address: Centro Nacional de Biotecnología (CNB), Campus de la Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain

5 Current address: Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2XY, UK

6 Current address: Department of Biochemistry, Sanger Building, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK

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Citation and License

Genome Biology 2004, 5:R84  doi:10.1186/gb-2004-5-10-r84

Published: 30 September 2004

Abstract

We have developed a systematic approach to generating cDNA clones containing full-length open reading frames (ORFs), exploiting knowledge of gene structure from genomic sequence. Each ORF was amplified by PCR from a pool of primary cDNAs, cloned and confirmed by sequencing. We obtained clones representing 70% of genes on human chromosome 22, whereas searching available cDNA clone collections found at best 48% from a single collection and 60% for all collections combined.