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通过荧光原位杂交检测CCND1基因座扩增

Detection of CCND1 Locus Amplification by Fluorescence In Situ Hybridization.

作者信息

Balázs Margit, Koroknai Viktória, Szász István, Ecsedi Szilvia

机构信息

Department of Preventive Medicine, Faculty of Public Health, University of Debrecen, Debrecen, Hungary.

MTA-DE Public Health Research Group, University of Debrecen, Debrecen, Hungary.

出版信息

Methods Mol Biol. 2018;1726:85-100. doi: 10.1007/978-1-4939-7565-5_9.

Abstract

It is well known that chromosomal aberrations of tumors are associated with the initiation and progression of malignancy. Fluorescence in situ hybridization (FISH) is a powerful, rapid method to detect chromosome copy number and structural alterations in tissue sections, chromosome, or interphase cellular preparations via hybridization of complementary probe sequences. The technique is based on the complementary nature of DNA double strands, which allows fluorescently labeled DNA probes to be used as probes to label the complementary sequences of target cells, chromosomes, and tissues. FISH technique has many applications, including basic gene mapping, used in pathological diagnosis to detect chromosome and gene copy number aberrations, translocations, microdeletions, and duplications. For the recognition of gene amplifications and deletions, locus-specific probes that are collections of one or a few cloned DNA sequences are routinely used. Multiplex-FISH (M-FISH) technique visualizes all chromosomes with different colors using spectrally distinct fluorophores for each chromosome in one experiment to detect numerical and structural alterations of chromosomes obtained from tumor cells. Recently many of the gene-specific probes are commercially available.

摘要

众所周知,肿瘤的染色体畸变与恶性肿瘤的发生和发展相关。荧光原位杂交(FISH)是一种强大、快速的方法,可通过互补探针序列的杂交来检测组织切片、染色体或间期细胞制剂中的染色体拷贝数和结构改变。该技术基于DNA双链的互补性质,这使得荧光标记的DNA探针能够用作探针来标记靶细胞、染色体和组织的互补序列。FISH技术有许多应用,包括基础基因图谱绘制,用于病理诊断以检测染色体和基因拷贝数畸变、易位、微缺失和重复。为了识别基因扩增和缺失,通常使用由一个或几个克隆DNA序列组成的位点特异性探针。多重荧光原位杂交(M-FISH)技术在一次实验中使用针对每条染色体的光谱不同的荧光团,用不同颜色使所有染色体可视化,以检测从肿瘤细胞获得的染色体的数量和结构改变。最近,许多基因特异性探针都有商业供应。

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