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Human Genome News Archive Edition

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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Human Genome Program, U.S. Department of Energy, Human Genome News (v6n4).

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