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The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.
Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.
Chromosome 19 Closure: High Resolution Physical Map of an Eight Megabase Region Extending from q12 to q13.1
Emilio Garcia, Jeff M. Elliott, Ann Gorvad, Linda K. Ashworth, Matt Burgin, Greg Lennon, Jane S. Lamerdin and Anthony V. Carrano
Human Genome Center, L-452, Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550.
We report the construction of a high-resolution physical map of an 8 Mb region that encompasses the entire q12 and most of the q13.1 bands of human chromosome l9. The ordered clone map has been obtained starting from a foundation of cosmid contigs assembled by automated fingerprinting and localized to the cytogenetic map by fluorescence in situ hybridization (FISH). Clonal continuity of the map has been achieved by binning and linking the pre-mapped cosmid contigs by means of YACs. The map consists of a single YAC contig with 108 YAC members (minimal spanning path of 14 YACs) linking 66 cosmid contigs. Eighteen STSs associated with genetic markers or derived from FISH-mapped cosmids have been placed on the map. Clonal continuity has been obtained by linking a series of pre-mapped cosmid contigs by means of YACs. In the first closure step, a series of YACs has been obtained by either STS-PCR screening of 75,000 YACs from the CEPH YAC library or by hybridization to a 800 member chromosome l9-specific MegaYAC sub library (Genethon) with Alu polymerase chain reaction (Alu-PCR) probes obtained from our ordered cosmid contigs. In a second step, the YACs obtained by this combined procedure have been used as templates to generate Alu-PCR probes against chromosome l9-specific cosmid arrays. This two step approach enables unambiguous identification of the cosmid-contig-YAC overlaps. Eighty percent (6 Mb) of the map obtained has been validated by Eco R I restriction mapping. This type of map provides a level of resolution an order of magnitude higher than those obtained from YACs alone.
This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under contract no. W-705-ENG-480.