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DNA microarray: a high throughput approach for methylation detection

机译:DNA芯片:用于甲基化检测的高通量方法

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We described a DNA microarray-based method combined with bisulphite treatment of DNA and regular PCR to examine hyper-methylation in promoter 1A of APC gene. A set of oligonucleotide probes were designed and immobilized on the aldehyde-coated glass slides for detecting the methylation pattern of 15 selected CpG sites in the region. The methylation status of 30 colorectal tumor samples have been examined by both of methylation-specific PCR (MS-PCR) and the present microarray method. The methylation pattern of the 15 CpG sites for the samples have been obtained with the microarray. A total of 19 samples out of 30 were methylated by microarray, in which five samples cannot be detected by MS-PCR due to the methylated CpG patterns not accordant to the MS-PCR primers. The detecting ratio for methylation of APC gene of colorectal tumor samples increased from 46.7% with MS-PCR to 63.3% with the microarray, which successfully demonstrated that DNA microarray-based method not only can obtained the methylation patterns for the related genes, but also decrease the false-negative results of methylation status by the conventional MS-PCR for the investigated genes. (c) 2004 Elsevier B.V. All rights reserved.
机译:我们描述了一种基于DNA微阵列的方法,结合亚硫酸氢盐处理DNA和常规PCR来检查APC基因启动子1A中的超甲基化。设计了一组寡核苷酸探针并将其固定在涂有醛的载玻片上,以检测该区域中15个选定CpG位点的甲基化模式。已经通过甲基化特异性PCR(MS-PCR)和当前的微阵列方法两者检查了30个结肠直肠肿瘤样品的甲基化状态。用微阵列已经获得了样品的15个CpG位点的甲基化模式。通过微阵列对30个样品中的19个样品进行了甲基化,其中5个样品由于甲基化的CpG模式与MS-PCR引物不一致而无法通过MS-PCR检测到。大肠肿瘤样品中APC基因甲基化的检出率由MS-PCR检测的46.7%提高到微阵列检测的63.3%,这成功地证明了基于DNA微阵列的方法不仅可以获得相关基因的甲基化模式,而且通过常规MS-PCR降低了所研究基因的甲基化状态的假阴性结果。 (c)2004 Elsevier B.V.保留所有权利。

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