果蝇睾丸中的低聚色素FISH。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-04-03 DOI:10.1007/7651_2025_613

Romir Raj, Vedansh Patel, Mayu Inaba

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引用次数: 0

摘要

荧光原位杂交（FISH）通常用于观察 RNA 和 DNA。它通常用于细胞遗传学和核型分析，检查染色体是否异常，诊断和鉴别疾病（Levsky 和 Singer，J Cell Sci 116:2833-2838, 2003）。Oligopaints 是一种新近建立的 DNA FISH 方法，因其灵活性和经济耐用性而迅速流行起来。寡核苷酸荧光染色法使用计算设计的可再生PCR寡核苷酸作为探针，可覆盖几千个碱基到整个染色体（Beliveau，Proc Natl Acad Sci 109：21301-21306，2012）。此外，不同的荧光团可用于多色成像所需的探针组。在这里，我们描述了一种优化的方法，用于实施寡染程序来观察果蝇睾丸中的基因组区域。我们进一步讨论了解析特定基因位点局部微观结构的可能性。

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Oligopaint FISH in Drosophila Testes.

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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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： 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.