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通过紫外线照射将聚(dT)修饰的DNA一步固定到用于微阵列的未修饰塑料基质上。

One-step immobilization of poly(dT)-modified DNA onto non-modified plastic substrates by UV irradiation for microarrays.

作者信息

Kimura Naoki

机构信息

Research and Development Center, Nisshinbo Industries Inc., 1-2-3 Onodai, Chiba-shi, Chiba 267-0056, Japan.

出版信息

Biochem Biophys Res Commun. 2006 Aug 25;347(2):477-84. doi: 10.1016/j.bbrc.2006.06.130. Epub 2006 Jun 30.

Abstract

Previously, 'DNattach', an alternative DNA immobilization system for attaching modified oligonucleotide probes onto a gold surface by UV irradiation that can be used in various DNA microarray applications including gene expression analysis, was developed. Attached to the gold surface, the modified probes have been shown to successfully detect synaptogenesis in the developing mouse cerebellum. In this study, this technology to immobilize modified oligonucleotide probes onto three different non-modified plastic surfaces in a microarray format has been further expanded. Using this system, single nucleotide polymorphism (SNP) genotyping of both oligonucleotide and PCR product targets has been successfully performed and it has also been shown that the probes immobilized on the slides can be used efficiently in hybridization experiments. Furthermore, it has been shown that probe concentrations of only 1-5 microM are sufficient for hybridization and that this immobilization method provides hybridization signals greater than those of conventional immobilization techniques.

摘要

此前,开发了“DNattach”,这是一种通过紫外线照射将修饰的寡核苷酸探针连接到金表面的替代DNA固定系统,可用于包括基因表达分析在内的各种DNA微阵列应用。连接到金表面的修饰探针已被证明能成功检测发育中小鼠小脑的突触形成。在本研究中,这项将修饰的寡核苷酸探针以微阵列形式固定在三种不同的未修饰塑料表面上的技术得到了进一步扩展。使用该系统,已成功对寡核苷酸和PCR产物靶标进行了单核苷酸多态性(SNP)基因分型,并且还表明固定在载玻片上的探针可有效地用于杂交实验。此外,已表明仅1-5 microM的探针浓度就足以进行杂交,并且这种固定方法提供的杂交信号比传统固定技术的信号更强。

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