Attachment of oligonucleotide probes to poly carbodiimide-coated glass for microarray applications
Attachment of oligonucleotide probes to poly carbodiimide-coated glass for microarray applications
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DOI：
10.1093/nar/gnh057
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发表时间：
2004-04-01
影响因子：
14.9
通讯作者：
Suzuki, O
中科院分区：
文献类型：
--
作者：
Kimura, N;Oda, R;Suzuki, O
基于寡核苷酸的DNA微阵列正成为分析基因表达和单核苷酸多态性（SNP）的越来越有用的工具。在这里，我们提出了一种方法，允许从未修饰的寡核苷酸的聚马来酰亚胺涂层的玻璃表面上通过紫外线照射的微阵列的制造。使用紫外线照射有利于信号强度水平的增加，但它不影响固定在表面上的寡核苷酸的信号辨别。使用未经修饰的寡核苷酸制备的阵列在紫外线照射下获得的信号强度比没有紫外线照射的信号强度高出7倍。SNPs的检测进行了测试，以确定这种技术是否可以区分特定的杂交信号，而不会造成显着的UV辐射诱导的损伤固定的寡核苷酸。我们发现，这种固定方法提供了更大的杂交信号和更好的匹配/错配比的SNPs比建立氨基硅烷技术。应用此技术制造DNA微阵列的序列分析进行了讨论。
Oligonucleotide-based DNA microarrays are becoming increasingly useful tools for the analysis of gene expression and single nucleotide polymorphisms (SNPs). Here, we present a method that permits the manufacture of microarrays from non-modified oligonucleotides on a poly carbodiimide-coated glass surface by UV-irradiation. The use of UV-irradiation facilitates an increase in the level of signal intensity, but it does not affect signal discrimination by the oligonucleotides immobilized on the surface. The signal intensity obtained for an array fabricated using non-modified oligonucleotides with UV-irradiation is similar to7-fold greater than that without UV-irradiation. The detection of SNPs was tested to ascertain whether this technique could discriminate specific hybridization signals without causing significant UV-irradiation-induced damage to the immobilized oligonucleotides. We found that this immobilization method provides greater hybridization signals and a better match/mismatch ratio of SNPs than do the established aminosilane techniques. Application of this technology to manufacturing DNA microarrays for sequence analysis is discussed.