Method
A genome annotation-driven approach to cloning the human ORFeome
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
Genome Biology 2004, 5:R84 doi:10.1186/gb-2004-5-10-r84
Published: 30 September 2004Abstract
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.
