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Direct immobilization of DNA probes on non-modified plastics by UV irradiation and integration in microfluidic devices for rapid bioassay

机译:通过紫外线照射将DNA探针直接固定在未改性的塑料上并整合到微流体设备中进行快速生物测定

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摘要

DNA microarrays have become one of the most powerful tools in the field of genomics and medical diagnosis. Recently, there has been increased interest in combining microfluidics with microarrays since this approach offers advantages in terms of portability, reduced analysis time, low consumption of reagents, and increased system integration. Polymers are widely used for microfluidic systems, but fabrication of microarrays on such materials often requires complicated chemical surface modifications, which hinders the integration of microarrays into microfluidic systems. In this paper, we demonstrate that simple UV irradiation can be used to directly immobilize poly(T)poly(C)-tagged DNA oligonucleotide probes on many different types of plastics without any surface modification. On average, five- and fourfold improvement in immobilization and hybridization efficiency have been achieved compared to surface-modified slides with aminated DNA probes. Moreover, the TC tag only costs 30% of the commonly used amino group modifications. Using this microarray fabrication technique, a portable cyclic olefin copolymer biochip containing eight individually addressable microfluidic channels was developed and used for rapid and parallel identification of Avian Influenza Virus by DNA hybridization. The one-step, cost-effective DNA-linking method on non-modified polymers significantly simplifies microarray fabrication procedures and permits great flexibility to plastic material selection, thus making it convenient to integrate microarrays into plastic microfluidic systems.Electronic supplementary materialThe online version of this article (doi:10.1007/s00216-011-5459-4) contains supplementary material, which is available to authorized users.
机译:DNA微阵列已成为基因组学和医学诊断领域最强大的工具之一。最近,人们越来越关注将微流控技术与微阵列相结合,因为这种方法在可移植性,减少分析时间,减少试剂消耗和增加系统集成方面具有优势。聚合物被广泛用于微流体系统,但是在这种材料上制造微阵列通常需要复杂的化学表面修饰,这阻碍了微阵列集成到微流体系统中。在本文中,我们证明了简单的紫外线照射可以直接将聚(T)聚(C)标签的DNA寡核苷酸探针固定在许多不同类型的塑料上,而无需进行任何表面修饰。与具有胺化DNA探针的表面修饰玻片相比,平均而言,固定化和杂交效率提高了五到四倍。而且,TC标签仅花费了常用氨基修饰的30%。使用这种微阵列制造技术,开发了包含八个可单独寻址的微流体通道的便携式环烯烃共聚物生物芯片,并将其用于通过DNA杂交快速,并行地鉴定禽流感病毒。在未修饰的聚合物上进行一步一步,经济高效的DNA链接方法可显着简化微阵列的制造程序,并为塑料材料的选择提供了极大的灵活性,从而使将微阵列集成到塑料微流体系统中变得十分方便。文章(doi:10.1007 / s00216-011-5459-4)包含补充材料,授权用户可以使用。

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