Abstract
This technique allows specific DNA sequences to be directly localized on chromosomes. The probe DNA is labeled by one of several methods that are described in this chapter. The labeled DNA is then applied onto chromosomes fixed on slides under conditions that allow specific hybridization of the probe to its complementary sequence on a chromosome. Following hybridization, unbound probe is removed by washing and the distribution of the probe visualized by autoradiography, immunofluorescence, or enzyme staining.
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
Buckle, V. J. and Craig, I. W. (1986) In situ hybridisation, in Human Genetic Diseases: A Practical Approach. Davies, K., ed. IRL, Oxford, pp. 85–100.
Zabel, B. U., Naylor, S. L., Sakaguchi, A. Y., Bell, G. I., and Shows, T. B. (1983) High resolution chromosomal localisation of human genes for amylase, proopiomelanocortin, somatostatin, and a DNA fragment (D351) by in situ hybridisation. Proc. Natl Acad Sci USA 80, 6932–6936.
Brown, R. M., Dahl, H.-H. M, and Brown, G. K. (1989) X-Chromosome localisation of the functional gene for the El alpha subunit of the human pyruvate dehydrogenase complex. Genomics 4, 174–181
Choo, K. H., Vissel, B, Brown, R, Filby, R. G., and Earle, E. (1988) Homologous alpha satellite sequences on human acrocentric chromosomes with selectivity for chromosomes 13,14 and 21: Implications for recombination between nonhomologues and Robertsonlan translocations. Nucleic Acids Res. 16, 1273–1284
Choo, K. H., Vissel, B., and Earle, E. (1989) Evolution of alpha satellite DNA on human acrocentric chromosomes Genomics 5, 332–344.
Brown, G. K., Brown, R. M., Scholem, R. D., Kirby, D M, and Dahl, H-H. M. (1989) The clinical and biochemical spectrum of human pyruvate dehydrogenase deficiency. Ann NY Acad. Sci. 573, 360–368.
Harper M E. and Saunders, G F (1981) Localisation of single copy DNAsequences on G-banded human chromosomes by in situ hybridisation Chromosoma 83, 431–439.
Choo, K H, Brown, R, Webb, G., Craig, I., and Filby, G. (1987) Genomic organisation of human centromenc alpha satellite DNA-Characterisation of a chromosome 17 alpha satellite sequence DNA 6, 297–305.
Choo, K H, Filby, G, Earle, E., and Brown, R. (1988) Isolation of human chromosome 21 sequences and their application to in situ hybridisation. Hum Genet 81, 49–53.
Albertson, D. G., Fishpool, R, Shernngton, P., Nacheva, E., and Milstein, C (1988) Sensitive and high resolution in situ hybridisation to human chromosomes using biotin labeled probes Assignment of the human thymocyte CD1 antigen genes to chromosome 1. EMBO J 7, 2801–2805
Bhatt, B, Burns, J, Flannery, D, and McGee, J O’D. (1988) Direct visualisation of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridisation. Nucleic Acids Res. 16, 3951–3961.
Chenf, D., Bernard, O, and Berger, R. (1989) Detection of single-copy genes by nonisotopic in situ hybridisation on human chromosomes Hum. Genet 81, 358–362.
Viegas-Pequignot, E, Dutnllaux, B, Magdelenat, H, and Coppey-Moisan, M (1989) Mapping of single-copy DNA sequences on human chromosomes by in situ hybridisation with biotinylated probes: Enhancement of detection sensitivity by intensified-fluorescence digital-imaging microscopy Proc Natl. Acad. Sci USA 86, 582–586
Burns, J, Chan, V T. W, Jonasson, J A., Fleming, K. A, Taylor, S, and McGee, J O’D (1985) Sensitivity system for visualising biotinylated DNA probes hybridised in situ. Rapid sex determination of intact cells. J. Clin Pathol. 38, 1085–1092.
Cremer, T, Landegent, J, Bruckner, A, Scholl, H. P., Schardin, M, Hager, H. D, Devilee, P., Pearson, P, and Van Der Ploeg, M. (1986) Detection of chromosome aberrations in the human mterphase nucleus by visualisation of specific target DNAs with radioactive and nonradioactive in situ hybridisation techniques: Diagnosis of trisomy 18 with probe LI 84 Hum Genet 74, 346–352.
Cremer, T, Lichter, P, Borden, J., Ward, D C, and Manuehdis, L (1988) Detecuon of chromosome aberrations in metaphase and mterphase tumor cells by in situ hybridisation using chromosome-specific library probes Hum. Genet. 80, 235–246
Lichter, P., Cremer, T., Tang, C. C, Watkins, P C, Manuehdis, L., and Ward, D. C (1988) Rapid detection of human chromosome 21 aberrations by in situ hybridisation Prvc NalL Acad. Sci. USA 85, 9664–9668.
Pinkel, D., Straume, T., and Gray, J W. (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridisation. Proc. Natl. Acad Sci USA 83, 2934–2938.
Lichter, P., Cremer, T., Borden, J., Manuelidis, L., and Ward, D. C. (1988) Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridisation using recombinant DNA libraries. Hum. Genet. 80, 224–234.
Webb, G. C., Lee, J. F., Campbell, H. D., and Young, I. G. (1989) Haemopoetic growth factor gene IL3 and IL4 mapped to the same locus on mouse chromosome 11 Cytogenet Cell Genet. 50, 107–110
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1991 The Humana Press Inc., Clifton, NJ
About this protocol
Cite this protocol
Choo, K.H., Brown, R.M., Earle, E. (1991). In Situ Hybridization of Chromosomes. In: Mathew, C.G. (eds) Protocols in Human Molecular Genetics. Methods in Molecular Biology, vol 9. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-205-1:233
Download citation
DOI: https://doi.org/10.1385/0-89603-205-1:233
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-205-7
Online ISBN: 978-1-59259-496-2
eBook Packages: Springer Protocols