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A flexible light-directed DNA chip synthesis gated by deprotection using solution photogenerated acids

机译:使用溶液光生酸通过脱保护进行门控的灵活的光导DNA芯片合成

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摘要

Oligonucleotide microarrays or oDNA chips are effective decoding and analytical tools for genomic sequences and are useful for a broad range of applications. Therefore, it is desirable to have synthesis methods of DNA chips that are highly flexible in sequence design and provide high quality and general adoptability. We report herein, DNA microarray synthesis based on a flexible biochip method. Our method simply uses photogenerated acid (PGA) in solution to trigger deprotection of the 5′-OH group in conventional nucleotide phosphoramidite monomers (i.e. PGA-gated deprotection), with the rest of the reactions in the synthesis cycle the same as those used for routine synthesis of oligonucleotides. The complete DNA chip synthesis process is accomplished on a regular DNA synthesizer that is coupled with a UV-VIS projection display unit for performing digital photolithography. Using this method, oDNA chips containing probes of newly discovered genes can be quickly and easily synthesized at high yields in a conventional laboratory setting. Furthermore, the PGA-gated chemistry should be applicable to microarray syntheses of a variety of combinatorial molecules, such as peptides and organic molecules.
机译:寡核苷酸微阵列或oDNA芯片是用于基因组序列的有效解码和分析工具,可用于广泛的应用。因此,期望有一种DNA芯片的合成方法,其在序列设计上高度灵活并且提供高质量和通用性。我们在本文中报道了基于柔性生物芯片方法的DNA微阵列合成。我们的方法仅使用溶液中的光生酸(PGA)来引发常规核苷酸亚磷酰胺单体中5'-OH基团的脱保护(即PGA门控脱保护),合成循环中的其余反应与用于寡核苷酸的常规合成。完整的DNA芯片合成过程是在常规DNA合成器上完成的,该合成器与用于执行数字光刻的UV-VIS投影显示单元耦合。使用这种方法,可以在常规实验室环境中快速轻松地合成含有新发现基因探针的oDNA芯片。此外,PGA门控化学应适用于各种组合分子(如肽和有机分子)的微阵列合成。

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