Introduction to the Workshop
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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 Specific Cosmid and Lambda Libraries from Flow-Sorted Chromosomes
Jeffrey A. Garnes, Benjamin S. Wong, Wanda Johnson, Jerry Eveleth, Anne Bergmann, Richard Langlois, Anthony V. Carrano, and Jeffrey C. Gingrich
Human Genome Center, L-452, Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550.
The National Laboratory Gene Library Project is responsible for the construction of human chromosome-specific cloned DNA libraries. The focus of phase II has been to construct partial MboI digest lambda (~10 kb) and cosmid (~40 kb) libraries for the entire human genome. The source of DNA for all of the libraries has been human chromosomes sorted from human/rodent hybrid somatic cell lines. Forty-five thousand cosmids cloned in the Lawrist 16 vector have been arrayed for chromosome 2 in addition to 23,000 fosmid clones. Approximately 63% of the clones in both libraries are derived from chromosome 2 and represent 7.5 chromosome equivalents. Approximately fifty thousand cosmids are being arrayed for chromosome 1 with additional DNA available for augmenting the library. The sort purity of this library as assessed by colony hybridization of a representative number of cosmids with both human and hamster DNA probes indicates that 61% of the sorted DNA is human. We are in the final stages of constructing chromosome-specific lambda libraries for chromosomes 1, 2 and 3. All of the lambda libraries will be deposited in the ATCC repository for distribution to the scientific community. This cooperative effort between Lawrence Livermore and Los Alamos National Laboratories has resulted in the production of large insert libraries for all of the human chromosomes. These libraries will continue to be invaluable resources in human genome mapping and analysis.
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.