Rapid High Resolution Single Nucleotide Polymorphism-Comparative Genome Hybridization Mapping in Caenorhabditis elegans
Rapid High Resolution Single Nucleotide Polymorphism-Comparative Genome Hybridization Mapping in Caenorhabditis elegans
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DOI：
10.1534/genetics.108.096487
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发表时间：
2009-01-01
期刊：
影响因子：
3.3
通讯作者：
Moerman, Donald G.
中科院分区：
文献类型：
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作者：
Flibotte, Stephane;Edgley, Mark L.;Moerman, Donald G.
我们已经开发了一种显着改进和简化的方法，用于秀丽隐杆线虫的表型性状的高分辨率映射使用单核苷酸多态性（SNP）和寡核苷酸阵列比较基因组杂交（阵列CGH）的组合。我们设计了一个定制的寡核苷酸阵列，使用典型野生型布里斯托菌株N2和夏威夷分离株CB 4856之间确认的SNP的子集，填充有对应于N2和CB 4856 SNP序列的密集重叠的50聚体探针。使用这种方法，在单个遗传杂交中，突变要求被定位到类似于200 kb的分辨率。6个突变分别代表C.使用来自少至100个突变N2/CB 4856杂合子的纯合突变分离子的群体的基因组DNA，明确地和高分辨率地检测了线虫染色体。我们的方法完全免除了标准SNP作图的PCR、限制性酶切消化和凝胶分析，应该很容易扩展到任何具有杂交菌株的生物体。这种方法将是特别强大的，当应用于困难或难以映射低的显着性表型。它也应该是可能的地图多基因性状使用这种方法。
We have developed a significantly improved and simplified method for high-resolution mapping of phenotypic traits in Caenorhabditis elegans using a combination of single nucleotide polymorphisms (SNPs) and oligo array comparative genome hybridization (array CGH). We designed a custom oligonucleotide array using a subset of confirmed SNPs between the canonical wild-type Bristol strain N2 and the Hawaiian isolate CB4856, populated with densely overlapping 50-mer probes corresponding to both N2 and CB4856 SNP sequences. Using this method a mutation call be mapped to a resolution of similar to 200 kb in a single genetic cross. Six mutations representing each of the C. elegans chromosomes were detected unambiguously and at high resolution using genomic DNA from populations derived from as few as 100 homozygous mutant segregants of mutant N2/CB4856 heterozygotes. Our method completely dispenses with the PCR, restriction digest, and gel analysis of standard SNP mapping and should be easy to extend to any organism with interbreeding strains. This method will be particularly powerful when applied to difficult or hard-to-map low-penetrance phenotypes. It should also be possible to map polygenic traits using this method.