{"title":"Comparison of Fluorescence in Situ Hybridization and Primed in Situ Labeling Methods for Detection of Single-Copy Genes in the Fungus Ustilago maydis","authors":"Shuxian Li, Charles P. Harris, Sally A. Leong","doi":"10.1006/emyc.1993.1028","DOIUrl":null,"url":null,"abstract":"<div><p>Li, S., Harris, C. P., and Leong, S. A. 1993. Comparison of fluorescence <em>in situ</em> hybridization and primed <em>in situ</em> labeling methods for detection of single-copy genes in the fungus <em>Ustilago maydis. Experimental Mycology</em> 17, 301-308. This report describes the use of fluorescence <em>in situ</em> hybridization for detection of single-copy genes in the fungus, <em>Ustilago maydis</em> . Either biotin- or digoxigenin-labeled DNA probes were hybridized to target nuclei of sporidia and subsequently rendered fluorescent by successive treatments with fluorescein (FlTC)-labeled avidin and biotinylated anti-avidin antibody or with anti-digoxigenin-FlTC or anti-digoxigenin-rhodamine. These results showed that the digoxigenin-based hybridization can be used successfully to specifically detect single-copy genes in nuclei of <em>U. maydis</em> with either 3.6- or 6.7-kb genomic DNA probes. By contrast, the biotin-based hybridization technique gave rise to nonspecific background. Primed <em>in situ</em> labeling with fluorescein-12-dUTP was also used successfully to specifically detect a 1.5-kb fragment of single-copy genomic DNA in nuclei of <em>U. maydis</em> sporidia. This technique has the advantages of lower staining background, shorter time of analysis, and a higher efficiency of hybridization of shorter DNA probes to target DNA, when compared to traditional <em>in situ</em> hybridization.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"17 4","pages":"Pages 301-308"},"PeriodicalIF":0.0000,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1993.1028","citationCount":"10","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental Mycology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0147597583710285","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 10
Abstract
Li, S., Harris, C. P., and Leong, S. A. 1993. Comparison of fluorescence in situ hybridization and primed in situ labeling methods for detection of single-copy genes in the fungus Ustilago maydis. Experimental Mycology 17, 301-308. This report describes the use of fluorescence in situ hybridization for detection of single-copy genes in the fungus, Ustilago maydis . Either biotin- or digoxigenin-labeled DNA probes were hybridized to target nuclei of sporidia and subsequently rendered fluorescent by successive treatments with fluorescein (FlTC)-labeled avidin and biotinylated anti-avidin antibody or with anti-digoxigenin-FlTC or anti-digoxigenin-rhodamine. These results showed that the digoxigenin-based hybridization can be used successfully to specifically detect single-copy genes in nuclei of U. maydis with either 3.6- or 6.7-kb genomic DNA probes. By contrast, the biotin-based hybridization technique gave rise to nonspecific background. Primed in situ labeling with fluorescein-12-dUTP was also used successfully to specifically detect a 1.5-kb fragment of single-copy genomic DNA in nuclei of U. maydis sporidia. This technique has the advantages of lower staining background, shorter time of analysis, and a higher efficiency of hybridization of shorter DNA probes to target DNA, when compared to traditional in situ hybridization.