Localizing genes in Drosophila melanogaster polytene chromosomes by fluorescence in situ hybridization |
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Authors: | Gavrila L Ecovoiu A A Georgescu L M |
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Affiliation: | Department of Genetics-CENTRAGEN, University of Bucharest, Bucharest, Romania |
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Abstract: | This paper describes a method for the identification of single copy genes in Drosophila melanogaster polytene chromosomes, using fluorescence in situ hybridization (FISH). We demonstrate the detection of white (w) , a gene previously mapped to 1-1.5 region of the linkage map, and to 3C2 region of the cytogenetic map of X chromosome. Squash preparations of polytene chromosomes from salivary glands dissected out from third instar larvae of Drosophila melanogaster were denatured and subjected to hybridization with a digoxigenin labeled probe, corresponding to mini-white gene. The preparations were then washed and incubated with antidigoxigenin-fluorescein antibodies. After removal of the nonspecifically bound antibodies, the polytene chromosomes were counterstained with propidium iodide. Fluorescence microscopy revealed white locus in the X chromosome in a subterminal location, in agreement with the above mentioned maps. The protocol is efficient and adaptable for simultaneously multiple signal detection. |
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Keywords: | Drosophila melanogaster polytene chromosomes white FISH labeled probe signal detection |
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