Abstract
Random fingerprinting strategies have been applied to a number of genome projects using unordered collections of phage or cosmid clones, e.g., Escherichia coli (1), Saccharomyces cerevisiae (2), and Caenorhabditis elegans (3). Overlaps were identified between either phage or cosmid clones using a fingerprinting method based on restriction endonuclease digestion. In each case the success of the method depended on the ability to generate and resolve sufficient numbers of bands in each fingerprint to detect as many as possible of the potential overlaps. Yeast artificial chromosomes (YACs) are not amenable to the same methods because of the presence of the yeast genome DNA in equimolar or higher amounts and because it is not possible to purify the artificial chromosome as it has no physical properties to distinguish it from the host DNA.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kohara, Y., Akiyaina, K., and Isono, K. (1987) The physical map of the whole E coli chromosome: application of a new strategy for rapid analysts and sorting of a large genomic library. Cell 50, 495–508.
Olson, M. V., Dutchik, J. E., Graham, M. Y., Brodeur, G. M., Helms, C., Frank, M., et al (1986) Random-clone strategy for genomic restriction mapping in yeast. Proc. Natl. Acad. Sci. USA 83, 7826–7830.
Coulson, A., Sulston, J., Brenner, S., and Karn, J. (1986) Towards a physical map of the genome of the nematode Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 83, 7821–7825.
Wada, M., Little, R. D., Abidi, F., Porta, G., Labella, T., Cooper, T., et al (1990) Human Xq24-Xq28. Approaches to mapping with yeast artificial chromosomes. Am. J. Hum. Genet. 46, 95–106.
Zucchi, I. and Schlessinger, D. (1992) Distribution of moderately repetitive sequences pTR5 and LF1 in Xq24-q28 human DNA and their use in assembling YAC contigs. Genomics 12, 264–275.
Nelson, D. L., Ledbetter, S. A., Corbo, L., Victoria, M. F., Ramirez-Solis, R., Webster, T. D., et al (1989) Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources. Proc. Natl. Acad. Sci. USA 86, 6686–6690.
Cole, C. G., Goodfellow, P. N., Bobrow, M., and Bentley, D. R. (1991) Generation of novel sequence tagged sites (STSs) from discrete chromosomal regions using Alu-PCR. Genomics 10, 816–826.
Coffey et al (1995) manuscript in preparation.
Coffey, A. J., Roberts, R. G., Green, E. D., Cole, C. G., Butler, R., Anand, R., et al. (1992) Construction of a 2 6 Mb contig in yeast artificial chromosomes spanning the human dystrophin gene using an STS-based approach. Genomics 12, 474–484.
Garoff, H. and Ansorge, W. (1981) Improvements of DNA sequencing gels. Anal. Biochem. 115, 450–457.
Cole, C. G., Dunham, I., Coffey, A. J., Ross, M. T., Meier-Ewert, S., Bobrow, M., and Bentley, D. R. (1992) A random STS strategy for construction of YAC contigs spanning defined chromosomal regions. Genomics 14, 256–262.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Humana Press Inc.
About this protocol
Cite this protocol
Coffey, A., Gregory, S., Cole, C.G. (1996). Alu-PCR Fingerprinting of YACs. In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:97
Download citation
DOI: https://doi.org/10.1385/0-89603-313-9:97
Publisher Name: Humana Press
Print ISBN: 978-0-89603-313-9
Online ISBN: 978-1-59259-541-9
eBook Packages: Springer Protocols