Abstract
Flow cytometry has provided the cytogeneticist with a fast and accurate method of measuring the quantity of DNA in each human chromosome (1). Almost all the chromosomes in the human complement can now be resolved and abnormal chromosomes and aneuploidies (13,21, and X) recognized. A flow karyotype shows a pattern of peaks and troughs that is unique for each individual or cell line because of the variation in heterochromatic regions of the chromosomes (2). When combined with family studies, flow cytometry has been able to resolve homologues differing in DNA content by as little as 1/2000 of the human genome (3,4), less than a metaphase band. In addition, the sorting capabilities of most flow machines have provided a method for the purification of small but useful quantities of individual chromosomes, for example, 2×106 average sized human chromosomes are equivalent to 500 ng of DNA. Using recombinant DNA techniques, this material can be used to generate a large number of DNA probes to produce a chromosome-specific library, which can be used for the molecular analysis of genetic disease (5,6). More recently, molecular biologists have experimented with gene mapping by sorting small quantities of individual chromosomes onto filters for spot-blot hybridization with DNA probes (7).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Langlois, R. G., Yu, L. C., Gray, J. W., and Carrano, A. V. (1982) Quantitative karyotyping of human chromosomes by dual beam flow cytometry. Proc. Natl. Acad. Sri. USA 79, 7876–7880.
Green, D. K., Fantes, J. A., Buckton, K. E., Elder, J. K., Malloy, P., Carothers, A., and Evans, H. J. (1984) Karyotyping and identification of human chromosome polymorphisms by single fluorochrome flow cytometry. Hum. Genet. 66, 143–146.
Harris, P., Boyd, E., Young, B. D., and Ferguson-Smith, M. A. (1986) Determination of the DNA content of human chromosomes by flow cytometry. Cytogenet. Cell Genet. 41, 14–21.
Harris, P., Cooke, A., Boyd, E., Young, B. D., and Ferguson-Smith, M. A. (1987) The potential of family flow karyotyping for the detection of chromosome abnormalities. Hum. Genet. 76, 129–133.
Krumlauf, R., Jeanpierre, M., and Young, B. D. (1982) Construction and characterisation of genomic libraries from specific human chromosomes. Proc. Natl. Acad. Sci. USA 79, 2971–2975.
Cooke, H. J., Fantes, J. A., and Green, D. K. (1983) Structure and evolution of human Y chromosome DNA. Differentiation 23, S48–S55.
Lebo, R. V., Gorin, F., Fletterick, R. J., Kao, F., Cheung, M., Bruce, B. D., and Kan, Y. W. (1984) High resolution chromosome sorting and DNA spot-blot analysis assign McArdle’s syndrome to chromosome 11. Science 225, 57–59.
Van de Sande, V. H., Lin, C. C., and Dengau, K. V. (1979) Clearly differentiated and stable chromosome bands produced by a spermine bis-acridine; a bifunctional analogue of quinacrine. Exp. Cell Res. 120, 439–444.
Sillars, R. and Young, B. D. (1981) A new method for the preparation of metaphase chromosomes for flow analysis. J. Histochem. Cytochem. 29, 74–78.
Melamed, M. R., Mullaney, P. F., and Mendelsohn, M. L. (eds.) (1979) Flow Cytometry and Sorting (Wiley, New York).
Van den Engh, G., Trask, B., and Gray, J. W. (1987) Improved resolution of bivariate flow karyotypes by manipulation of staining conditions. Cytometry Suppl. 1, 3.
Porteous, D. J., Morten, J. E. N., Cranston, G., Fletcher, J. M., Mitchell, A., van Heyningen, V., Fantes, J. A., Boyd, P. A., and Hastie, N. D. (1986) Molecular and physical arrangements of human DNA in HRAS1-selected, chromosome mediated transfectants. Mol. Cellular Biol. 6, 2223–2232.
Trask, B., Van den Engh, G., Gray, J. W., Vanderlaan, M., and Turner, B. (1984) Immunofluorescent detection of histone 2B on metaphase chromosomes using flow cytometry. Chromosoma 90, 295–302.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1990 The Humana Press Inc.
About this protocol
Cite this protocol
Fantes, J.A., Green, D.K. (1990). Human Chromosome Analysis and Sorting. In: Walker, J.M., Pollard, J.W., Walker, J.M. (eds) Animal Cell Culture. Methods in Molecular Biology, vol 5. Humana Press. https://doi.org/10.1385/0-89603-150-0:529
Download citation
DOI: https://doi.org/10.1385/0-89603-150-0:529
Publisher Name: Humana Press
Print ISBN: 978-0-89603-150-0
Online ISBN: 978-1-59259-492-4
eBook Packages: Springer Protocols