{"title":"Oligopaint FISH in Drosophila Testes.","authors":"Romir Raj, Vedansh Patel, Mayu Inaba","doi":"10.1007/7651_2025_613","DOIUrl":null,"url":null,"abstract":"<p><p>Fluorescence in situ hybridization (FISH) is commonly performed to visualize RNA and DNA. It is routinely used in cytogenetics and karyotyping to check for chromosomal abnormalities and diagnose and identify diseases (Levsky and Singer, J Cell Sci 116:2833-2838, 2003). Oligopaints is a recently established DNA FISH method that has quickly become popular because of its flexibility and economical durability. Oligopaints uses computationally designed PCR-renewable oligonucleotides for probes, which can cover a few kilobases to whole chromosomes (Beliveau, Proc Natl Acad Sci 109:21301-21306, 2012). In addition, different fluorophores can be used for desired probe sets for multicolor imaging. Here, we describe an optimized method for implementing the Oligopaints procedure to visualize genomic regions in Drosophila testis. We further discuss the possibility of resolving local microstructure of specific gene loci.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/7651_2025_613","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
Fluorescence in situ hybridization (FISH) is commonly performed to visualize RNA and DNA. It is routinely used in cytogenetics and karyotyping to check for chromosomal abnormalities and diagnose and identify diseases (Levsky and Singer, J Cell Sci 116:2833-2838, 2003). Oligopaints is a recently established DNA FISH method that has quickly become popular because of its flexibility and economical durability. Oligopaints uses computationally designed PCR-renewable oligonucleotides for probes, which can cover a few kilobases to whole chromosomes (Beliveau, Proc Natl Acad Sci 109:21301-21306, 2012). In addition, different fluorophores can be used for desired probe sets for multicolor imaging. Here, we describe an optimized method for implementing the Oligopaints procedure to visualize genomic regions in Drosophila testis. We further discuss the possibility of resolving local microstructure of specific gene loci.
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For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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