A combinatorial approach for fast, high-resolution mapping
A combinatorial approach for fast, high-resolution mapping
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DOI：
10.1006/geno.2002.6809
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发表时间：
2002-08-01
期刊：
影响因子：
4.4
通讯作者：
Bensimon, A
中科院分区：
文献类型：
--
作者：
Conti, C;Bensimon, A
高分辨率物理图谱可用作后续多项研究的支架，例如测序项目以及疾病基因和调节基因表达的遗传元件的定位克隆。在这里，我们描述了一种基于组合多重 FISH 方法对拉伸 DNA 分子进行快速、高分辨率物理绘图的方法。荧光标记被分配给二进制代码，探针根据其标记通过二进制标签进行识别。为了验证该方法，我们通过三种杂交测定绘制了覆盖人类 11 号染色体上约 300 kb 区域的 8 个探针。这种方法使人们能够通过其克隆的顺序来确定大区域的结构组织，而不会产生歧义。对照细胞中建立的结构为进一步研究提供参考，以检测疾病细胞表现出的重排并发现不同细胞类型和生物体表现出的差异。
High-resolution physical maps can be used as a scaffold for several subsequent studies, such as sequencing projects and positional cloning of disease genes and genetic elements that regulate gene expression. Here we describe a method for fast, high-resolution physical mapping on stretched DNA molecules, based on a combinatorial multi-FISH approach. Fluorescent labels are assigned to a binary code and probes are identified by a binary tag according to their labeling. To validate the approach, we have mapped eight probes covering a region of about 300 kb on human chromosome 11 with three hybridization assays. This approach enables one to determine the structural organization of a large region by means of the order of its clones, without ambiguities. The structure established in a control cell constitutes a reference for further studies, to detect rearrangements displayed by disease cells and to find differences shown by different cell types and organisms.