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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 the Entire q12 and q13.1 Bands

Jeffrey Elliott[1], Ann Gorvad[1], K. Soliman[2], Brian Cheney[1], Linda K. Ashworth[1], Matt Burgin[1], Jane S. Lamerdin[1], Greg Lennon[1], Anthony V. Carrano[1], and Emilio Garcia[1,3]
[1]Human Genome Center, L-452, Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550. [2]Department of Plant and Soil Science, A&M University, Huntsville, Alabama. [3]Corresponding author.

A 12.5 Mb, high-resolution map of a region that encompasses the entire q12 and q13.1 bands of human chromosome 19 has been constructed. 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) [1,2]. 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 comprised of 145 YAC members (minimal spanning path of 18 YACs) linking more than 100 cosmid contigs. Nineteen 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 [3,4] or by hybridization to a 800 member chromosome 19-specific Mega YAC sub library (Genethon) [5] with Alu polymerase chain reaction (Alu-PCR) probes obtained from our 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 19-specific cosmid arrays. This two step approach enables unambiguous identification of the cosmid-contig-YAC overlaps. Seventy seven percent (9531 Kb) of the map obtained has been validated by EcoRI restriction mapping. This type of map provides a level of resolution and 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-7405-ENG-48.

[1] Brandriff, B.F., Gordon, L.A., Fertitta, A., et al (1994) Genomics. In Press.
[2] Trask, et al (1993) Genomics, 15, 133-145.
[3] Albertson, et al (1990) PNAS, 87, 4526.
[4] Dausset, et al (1992) Behring Inst. Mitt, 91, 13-20.
[5] Chumakov, et al (1992) Nature Genetics, 1, 222-225.