Ordering STSs via YAC hybridization to radiation hybrids using Inner Product Mapping
- Carnegie Mellon Univ., Pittsburgh, PA (United States)
- Univ. of Pittsburgh, PA (United States)
We have developed a general system for rapid construction and integration of long-range, high resolution physical and transcription maps. To demonstrate this approach, a complete 4.5 Mbp YAC clone contig encompassing D11S12 to cathepsin D (CTSD) on human chromosome 11p15 has been assembled from the 1991 CEPH YAC library, the CEPH MEGA YAC library, and the Show`s chromosome 11-specific YAC library. All three libraries have been arranged for efficient, high throughput screening by STS content, an interpreted repeat element (IRE)-PCR-based filter hybridization scheme, and single colony hybridization filters. This YAC contig shows significant overlap, is anchored by a large number of established genetic loci and orders over 250 new markers. A complete cosmid contig spanning this region has also been assembled. Complex hybridization probes, corresponding to YACs spanning the contig, were developed by IRE-bubble PCR and used to screen high density colony grids representing an arrayed, flow-sorted chromosome 11 cosmid library. Once identified and ordered, these cosmids have been processed through an exon amplification scheme to identify transcribed sequences which are specific to this region. On average, we have identified one unique exon for every 12.5 kb of genomic sequence across the entire 4.5 Mbp region. These exons have in turn been ordered and mapped within the context of our high resolution physical map, YAC contig and cosmid contig. Finally, we are in the process of isolating cDNAs specific to these exons using a set of arrayed cDNA libraries which have been arranged into a complex set of pools for rapid and efficient screening by PCR on hybridization.
- OSTI ID:
- 134449
- Report Number(s):
- CONF-941009--
- Journal Information:
- American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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