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

摘要

We have developed a general method foren masselsolation of cDNAs present in a normalized library by hybridization to arrayed chromosome-specific phageλclones; we have used this approach to initiate exon-mapping of human chromosome 13. An advantage of the simultaneous isolation of cDNA/λpairs is that it allows cytogenetic assignment of a bona fide genomic clone byin situhybdridization, 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 func