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© 1994 Oxford University Press

OTHER

Selection of cDNAs using chromosome-specific genomic clones: application to human chromosome 13

Maria de Fatima1,4, Ming-Tsung Yu2, Plerre Jelenc1,4, Stephen Brown3, Long Su1,4, Lee Lawton1,4, Larry Deaven5, Argiris Eistratiadis2, Dorothy Warburton2 and Marcelo Bento Soares1,4,*

1Department of Psychiatry, Columbia University New York, NY 10032 2Department of Genetics and Development, Columbia University New York, NY 10032 3Department of Obstetrics and Gynecology, Columbia University New York, NY 10032 4The New York State Psychiatric Institute New York, NY 10032 5Center for Human Genome Studies, Los Alamos National Laboratory Los Alamos, NM 87545, USA

*To whom correspondence should be addressed at: Department of psychatry of Columbia University and The New York State Psychatric Institute, 772 West 168th Street, Unit #41, New York, NY 10032, USA

Received June 10, 1994; Revised June 27, 1994; Accepted June 27, 1994

We have developed a general method for en masse lsolation of cDNAs present in a normalized library by hybridization to arrayed chromosome-specific phage {lambda} clones; we have used this approach to initiate exon-mapping of human chromosome 13. An advantage of the simultaneous isolation of cDNA/{lambda} pairs is that it allows cytogenetic assignment of a bona fide genomic clone by in situ hybdridization, which also verifies that the corresponding cDNA or a homologous expressed sequence resides on chromosome 13. This information is enriched by partial sequencing of a selected cDNA from both ends. The sequence of the 3' noncoding region provides an ‘Identifier’ that is used to develop STSs, while the sequence from the 5' end, often corresponding to a coding region, is used for homology searches in databases that occasionally reveal gene functions.


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