Generating a physical map of chromosome 11
- Roswell Park Cancer Institute, Buffalo, NY (United States); and others
A four-hit chromosome-specific YAC library is being used to construct an overlapping YAC clone physical map of human chromosome 11. Each of the 1824 YAC clones have been sized by pulsed-field gel analysis with a resulting average insert size of 350 kb. Three hundred of the clones have been localized to a chromosome band by fluorescence in situ hybridization (FISH). We are using two complementary approaches to assemble YAC contigs: hybridization-based screening with Alu-PCR products and STS-based screening. Both screening approaches use pooling strategies to uniquely address each of the well locations with a minimum number of DNA pools. Since our main source of YAC clones is a chromosome specific library, the pool complexity is an order of magnitude smaller in comparison to total genomic libraries. Alu products with three different Alu primers were generated from both the monochromosomal J1 hybrid and individual YAC clones. YAC clones were chosen to generate probes using a sample without replacement strategy. Greater than 1700 YAC clones have been assembled into contigs. These initial contigs are estimated to be on average 1 Mb in size. Each of the contigs have been binned into a chromosomal band by virture of it containing YACs localized by FISH or YACs positive for one or more mapped STSs. Confirmation of the contigs is being achieved with Line I fingerprinting. The initial contigs are currently being joined using probes derived from the YAC clone ends defining the outermost points in the contigs for hybridization of colony filters. Remaining gaps in the contigs will be joined using alternative large insert libraries.
- OSTI ID:
- 134454
- 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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