Fluorescence In Situ Hybridization onto DNA Fibres Generated Using Molecular Combing
Fluorescence In Situ Hybridization onto DNA Fibres Generated Using Molecular Combing
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DOI：
10.1007/978-3-662-52959-1_31
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发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Yang, Fengtang
中科院分区：
文献类型：
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作者：
Louzada, Sandra;Komatsu, Jun;Yang, Fengtang
分子精梳是制备延伸DNA纤维的一种先进方法。它使数百个DNA分子在玻璃表面上均匀平行地伸展，以每微米2千碱基对的前所未有的分辨率。当与荧光原位杂交（FISH）相结合时，分子梳理可以直接可视化基因组结构和拷贝数变化，以及量化基因组组装中序列contigs之间重叠和间隙的大小。在这里，我们提出了一种多色纤维- fish方案，使用分子梳理制备的DNA纤维。
Molecular combing represents an advanced method for preparing extended DNA fibres. It enables uniform and parallel stretching of hundreds of DNA molecules on a glass surface, at an unprecedented resolution of two kilobase pairs per micrometre. When coupled with fluorescence in situ hybridization (FISH), molecular combing allows the direct visualisation of genomic structure and copy number variation, as well as the quantification of sizes of overlap and gap between sequence contigs in the genome assembly. Here we present a multicolor fibre-FISH protocol using DNA fibres prepared by molecular combing.