Sponsored by the U.S. Department of Energy Human Genome Program
Human Genome News, Nov. 1994; 6(4):12
Washington University School of Medicine
WASHINGTON UNIVERSITY SCHOOL OF MEDICINE (WUSM)
- (NIH, established 1990)
- DAVID SCHLESSINGER, Director
- Bernard H. Brownstein, Assistant Director
- CONTACT: Brownstein (314/362-3613 or -1199, Fax: -3203, buddy@sequencer.wustl.edu); Center for Genetics in Medicine; 4566 Scott Ave., Box 8232; St. Louis, MO 63110.
OTHER KEY RESEARCHERS
- Frank Burough
- Ellson Chen (Applied Biosystems Division, Perkin-Elmer)
- Terry Featherstone
- Pui Kwok
- Ramaiah Nagaraja
- Volker Nowotny
- John Rice
- David States
MAJOR GOALS
- Construction of sequence tagged site-based integrated physical and genetic maps of the X chromosome.
- Implementation of megabase-level sequencing of selected X chromosome regions.
MAJOR ACCOMPLISHMENTS
- Completion (with collaborators) of yeast artificial chromosome coverage of a number of loci, including 2 Mb in the Huntington's disease region and 4 Mb in the major histocompatibility complex.
- Formulation, testing, and implementation of sequence tagged site-content mapping.
- Assembly of contigs >65% of chromosome 7 from materials that include >147 sequence tagged sites [of which 100 are highly polymorphic linkage probes obtained originally from Jean Weissenbach (Institut Pasteur) and Ray White (Univ. of Utah)] and a set of >5400 chromosome 7-specific yeast artificial chromosomes that are low in cocloning.
- Assembly of over 85% of the X chromosome in contigs using 1400 sequence tagged sites and 5000 X-specific yeast artificial chromosomes. The contigs range up to 21 Mb in length, with fully rationalized maps available for Xq26-qter. Large contigs are now being aligned in Xp, Xq13, and Xq24-q26; 200 highly polymorphic gene-specific markers are being placed along the chromosome with cognate yeast artificial chromosomes.
- Sequence tagged site and yeast artificial chromosome information has been submitted to Genome Data Base.
- Wide distribution of Wash. Univ. human yeast artificial chromosome library. Submission to American Type Culture Collection for worldwide distribution of all X chromosome-specific yeast artificial chromosomes, including an X-specific yeast artificial chromosome library.
- Assembly of robot-assisted workstation to screen yeast artificial chromosome libraries; includes capacity for up to 1800 polymerase chain reaction reactions/d.
- Development of algorithms and software for analysis of sequence tagged site-content and radiation hybrid mapping data and for choosing polymerase chain reaction primers.
- Design and construction of a database for sequence tagged site-content mapping data.
- Implementation of SEGMAP for visual map representation and for testing sequence tagged site-based DNA segment contigs.
AVAILABLE RESOURCES
- Assorted genomic and X-specific yeast artificial chromosome libraries (made at Wash. Univ. and elsewhere); all configured for polymerase chain reaction-screening yeast artificial chromosomes with sequence tagged sites.
- Implemented rapid polymerase chain reaction screening of X-specific cosmid libraries.
- High-throughput polymerase chain reaction machine.
- Information on polymerase chain reaction conditions and end-cloning protocols.
- UNIX-based software to run a Zymark side-loader arm in conjunction with a Biomek 1000 robot for robot-assisted screening, and information for preparing large libraries for polymerase chain reaction screening.
- SEGMAP (yeast artificial chromosome contig assembly software).
- Reliable procedures to recover yeast artificial chromosome insert ends and sequence them automatically.
- Software for sequence tagged site development and data storage.
HGMIS staff
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The electronic form of the newsletter may be cited in the following style:
Human Genome Program, U.S. Department of Energy, Human Genome News (v6n4).
