Identification of coding single nucleotide polymorphisms and mutations by combination of genome tiling arrays and enrichment/depletion of mismatch cDNAs.
Identification of coding single nucleotide polymorphisms and mutations by combination of genome tiling arrays and enrichment/depletion of mismatch cDNAs.
复制标题
通过组合基因组平铺阵列和错配 cDNA 的富集/去除来鉴定编码单核苷酸多态性和突变。
DOI：
10.1016/j.ab.2006.05.013
复制
发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Tang,Ling
中科院分区：
文献类型：
--
作者：
Liu,Meng-Min;Weissman,ShermanM;Tang,Ling
基因组拼接阵列技术结合富集和去除含有错配的cDNA片段的方法，为检测合并cDNA样品中的编码单核苷酸多态性（cSNPs）和突变提供了有用的方法。通过使用错配修复胸腺嘧啶DNA糖基化酶结合的珠粒进行选择，从人原发性黑色素瘤细胞和正常黑色素细胞中获得富集的错配和完全匹配的cDNA样品。然后标记这些cDNA样品并与DNA元件百科全书基因组拼接阵列杂交。结果显示，富集的错配样本的潜在cDNA变异区域的杂交强度值增加，而富集的完全匹配样本的相应区域的杂交强度值降低。通过聚合酶链反应产物测序证实了六种潜在的突变，包括黑色素瘤细胞中的两种新的杂合突变。我们认为，这种策略应提高整个人类基因组的cSNP和突变检测的效率，并降低cDNA变异的全基因组分析的成本和复杂性。
Genome tiling array technology combined with a method for both enrichment and depletion of mismatch-containing cDNA fragments offers a useful approach for detecting coding single nucleotide polymorphisms (cSNPs) and mutations in pooled cDNA samples. Enriched mismatch and perfect match cDNA samples from human primary melanoma cells and normal melanocytes were obtained by selection using mismatch repair thymine DNA glycosylase-bound beads. These cDNA samples were then labeled and hybridized to Encyclopedia of DNA Elements genome tiling arrays. The results revealed that the hybridization intensity values of potential cDNA variation regions of the enriched mismatch samples increased, whereas the hybridization intensity values of corresponding regions of the enriched perfect match samples decreased. Six potential mutations were confirmed by polymerase chain reaction product sequencing, including two novel heterozygous mutations in melanoma cells. We suggest that this strategy should increase the efficiency of both cSNP and mutation detection throughout the entire human genome and decrease the cost and complexity of genomewide analysis of cDNA variations.