A High-Resolution Physical Map of Human Chromosome 11

The development of a highly reliable physical map with landmark sites spaced an average of 100 kbp apart has been a central goal of the Human Genome Project. We have approached the physical mapping of human chromosome 11 with this goal as a primary target. We have focused on strategies that would ut...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1996-04, Vol.93 (7), p.3149-3154
Main Authors: Qin, Shizhen, Nowak, Norma J., Zhang, Jialu, Sheila N. J. Sait, Mayers, Peter G., Higgins, Michael J., Cheng, Yi-Jun, Li Li, Munroe, David J., Gerhard, Daniela S., Weber, Bernhard H., Bric, Eva, Housman, David E., Evans, Glen A., Shows, Thomas B.
Format: Article
Language:English
Subjects:
Chimera
Chromosome Mapping
Chromosomes
Chromosomes, Artificial, Yeast
Chromosomes, Human, Pair 11
Cloning, Molecular
DNA
DNA - chemistry
DNA probes
Gels
Genetic screening
Genetics
Genomes
Genomics
Human Genome Project
Humans
In Situ Hybridization, Fluorescence
Libraries
Polymerase Chain Reaction
Restriction Mapping
Sequence tagged sites
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Summary:The development of a highly reliable physical map with landmark sites spaced an average of 100 kbp apart has been a central goal of the Human Genome Project. We have approached the physical mapping of human chromosome 11 with this goal as a primary target. We have focused on strategies that would utilize yeast artificial chromosome (YAC) technology, thus permitting long-range coverage of hundreds of kilobases of genomic DNA, yet we sought to minimize the ambiguities inherent in the use of this technology, particularly the occurrence of chimeric genomic DNA clones. This was achieved through the development of a chromosome 11-specific YAC library from a human somatic cell hybrid line that has retained chromosome 11 as its sole human component. To maximize the efficiency of YAC contig assembly and extension, we have employed an Alu-PCR-based hybridization screening system. This system eliminates many of the more costly and time-consuming steps associated with sequence tagged site content mapping such as sequencing, primer production, and hierarchical screening, resulting in greater efficiency with increased throughput and reduced cost. Using these approaches, we have achieved YAC coverage for >90% of human chromosome 11, with an average intermarker distance of
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.93.7.3149