A genome annotation-driven approach to cloning the human ORFeome

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 repres...

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Bibliographic Details
Published inGenome biology Vol. 5; no. 10; p. R84
Main Authors Collins, John E, Wright, Charmain L, Edwards, Carol A, Davis, Matthew P, Grinham, James A, Cole, Charlotte G, Goward, Melanie E, Aguado, Begoña, Mallya, Meera, Mokrab, Younes, Huckle, Elizabeth J, Beare, David M, Dunham, Ian
Format Journal Article
LanguageEnglish
Published England BioMed Central Ltd 01.01.2004
BioMed Central
Subjects
Chromosomes, Human, Pair 22 - genetics
Cloning, Molecular - methods
Computational Biology
Databases, Genetic
DNA, Complementary - genetics
Genome, Human
Genomics - methods
Humans
Method
Open Reading Frames - genetics
Polymerase Chain Reaction
Polymorphism, Single Nucleotide - genetics
Proteome - genetics
Research Design
Sequence Analysis, DNA
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Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:1474-760X
1465-6906
1474-760X
1465-6914
DOI:10.1186/gb-2004-5-10-r84