Abstract
A non-radioactive method for in situ hybridization of cosmid probes to metaphase chromosomes is described. Two procedures are involved: (i) hybridization with a cosmid probe labelled by nick translation in the presence of digoxigenin dUTP. The signal is visualized by an alkaline phosphatase conjugated antibody. (ii) FPG banding of the chromosomes. The steps involved in these two procedures are combined in an order which allows simultaneous observation of the banding pattern and the hybridization signal. The metaphases can thus be analysed after a single photographic step. This technique is considerably simpler than the method used previously.
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References
Dutrillaux B, Lejeune J (1971) Sur une nouvelle technique d'analyse du caryotype humain. CR Acad Sci Paris 272:2638
Garson JA, van den Berghe J, Kemshead JT (1987) Novel nonisotopic in situ hybridization technique detects small (1 Kb) unique sequences in routinely G-banded human chromosomes: fine mapping of N-myc andβ-NGF genes. Nucleic Acids Res 15:4761–4769
Harper ME, Ullrich A, Saunders GF (1981) Localization of the human insulin gene to the distal end of the short arm of chromosome 11. Proc Natl Acad Sci USA 78:4458–4460
Langer-Safer PR, Levine M, Ward DC (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci USA 79:4381–4385
Little PFR (1987) Choice and use of cosmid vectors. In: Glover DM (ed) Gene cloning, vol 3. IRL Press, Oxford, pp 19–42
Mattei MG, Philip N, Passage E, Moisan JP, Mandel JL, Mattei JF (1985) DNA probe localization at 18p113 band by in situ hybridization and identification of a small supernumerary chromosome. Hum Genet 69:268–271
Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934–2938
Viegas-Péquignot E, Dutrillaux B, Magdelenat H, Coppey-Moisan M (1989) Mapping of single-copy DNA sequences on human chromosomes by in situ hybridization with biotinylated probes: Enhancement of detection sensitivity by intensified-fluorescence digital-imaging microscopy. Proc Natl Acad Sci USA 85:582–586
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Zhang, F.R., Heilig, R., Thomas, G. et al. A one-step efficient and specific non-radioactive non-fluorescent method for in situ hybridization of banded chromosomes. Chromosoma 99, 436–439 (1990). https://doi.org/10.1007/BF01726696
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DOI: https://doi.org/10.1007/BF01726696