1. A genome annotation-driven approach to cloning the human ORFeome.
- Author
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Collins JE, Wright CL, Edwards CA, Davis MP, Grinham JA, Cole CG, Goward ME, Aguado B, Mallya M, Mokrab Y, Huckle EJ, Beare DM, and Dunham I
- Subjects
- Chromosomes, Human, Pair 22 genetics, Computational Biology, DNA, Complementary genetics, Databases, Genetic, Humans, Polymerase Chain Reaction, Polymorphism, Single Nucleotide genetics, Research Design, Sequence Analysis, DNA, Cloning, Molecular methods, Genome, Human, Genomics methods, Open Reading Frames genetics, Proteome genetics
- 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.
- Published
- 2004
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