Abstract
The ordering of yeast artificial chromosome (YAC) clones by sequence-tagged site (STS)-content mapping has proven an effective means of constructing large, contiguously cloned arrays of DNA, some of which span almost entire human chromosomes (1). This method requires that each YAC clone be tested for the presence or absence of each STS in a simple binary fashion and ultimately results in a content map where each clone is identified and positioned relative to others based on its complement of STSs (2,3). The application of this technique to the long-range physical mapping of the genomes of more complex organisms, although powerful, is complicated by the fact that precise distance and positional information cannot be derived directly from the STS content of each clone.
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References
Foote, S., Vollrath, D., Hilton, A., and Page, D. C. (1992) The human Y chromosome, overlapping DNA clones spanning the euchromatic region. Science 258, 60–66.
Green, E. D. and Green, P. (1991) Sequence-tagged site (STS) content mapping of human chromosomes: theoretical considerations and early experiences. PCR Methods Appl. 1, 77–90.
Green, E. D. and Olson, M. V. (1990) Chromosomal region of the cystic fibrosis gene in yeast artificial chromosomes a model for human genome mapping. Science 250, 94–98.
Ferrin, L. J. and Camerini-Otero, R. D. (1991) Selective cleavage of human DNA: RecA-assisted restriction endonuclease (RARE) cleavage. Science 254, 1494–1497.
Koob, M., Burkiewicz, A., Kur, J., and Szybalski, W. (1992) RecA-AC single-site cleavage of plasmids and chromosomes at any predetermined restriction site. Nucleic Acids Res. 20, 5831–5836.
Gnirke, A., Barnes, T. S., Patterson, D., Schild, D., Featherstone, T., and Olson, M. V. (1991) Cloning and in vivo expression of the human GART gene using yeast artificial chromosomes. EMBO J., 10, 1629–1634.
Gnirke, A., Iadonato, S. P., Kwok, P. Y., and Olson, M. V. (1994) Physical calibration of yeast artificial chromosome contig maps by RecA-assisted restriction endonuclease (RARE) cleavage. Genomics 24, 199–210.
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© 1996 Humana Press Inc.
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Iadonato, S.P., Gnirke, A. (1996). RARE-Cleavage Analysis of YACs. In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:75
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DOI: https://doi.org/10.1385/0-89603-313-9:75
Publisher Name: Humana Press
Print ISBN: 978-0-89603-313-9
Online ISBN: 978-1-59259-541-9
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