Programmable Chromosome Painting with Oligopaints.
Programmable Chromosome Painting with Oligopaints.
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使用 Oligopaints 进行可编程染色体绘画。
DOI：
10.1007/978-1-4939-9674-2_11
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Joyce,EricF
中科院分区：
文献类型：
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作者：
Nguyen,SonC;Joyce,EricF
目前通过荧光原位杂交(FISH)绘制染色体的方法成本高、耗时长，而且复杂性有限。与传统的探针源不同，寡聚体是通过计算设计的，在微阵列上合成，并通过聚合酶链式反应进行扩增。这种方法允许精确控制它们的目标序列，范围从几千个碱基到整个染色体，使用相同的基本方案。我们利用齐聚颜料的灵活性和可扩展性，为果蝇、小鼠和人类染色体生成了低成本和可再生的染色体颜料。这些寡色素库可以定制为以染色体范围的方式标记任何基因组特征(S)。此外，这种方法与序列FISH兼容，只需一个变性步骤就可以标记整个基因组。在这里，我们概述了一个协议和考虑，以扩大寡不显色技术的荧光标记整个染色体。
Current methods for chromosome painting via fluorescence in situ hybridization (FISH) are costly, time-consuming, and limited in complexity. In contrast to conventional sources of probe, Oligopaints are computationally designed, synthesized on microarrays, and amplified by PCR. This approach allows for precise control over the sequences they target, which can range from a few kilobases to entire chromosomes with the same basic protocol. We have utilized the flexibility and scalability of Oligopaints to generate low-cost and renewable chromosome paints for Drosophila, mouse, and human chromosomes. These Oligopaint libraries can be customized to label any genomic feature(s) in a chromosome-wide manner. Additionally, this method is compatible with sequential FISH to label entire genomes with a single denaturation step. Here, we outline a protocol and considerations to scale the Oligopaint technology for fluorescent labeling of whole chromosomes.