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A scalable method for multiplex LED-controlled synthesis of DNA in capillaries

机译:用于毛细管中DNA的LED多重控制合成的可扩展方法

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摘要

As research in synthetic biology and genomic sciences becomes more widespread, the need for diverse oligonucleotide populations has increased. To limit reagent cost, it would be advantageous to obtain high quality populations in minute amounts. Towards that end, synthesis of DNA strands in capillaries utilizing photolabile 3-nitrophenylpropyloxycarbonyl (NPPOC) chemistry and ultraviolet-light emitting diodes (UV-LEDs) was examined. Multiple oligonucleotides were made in single capillaries and were characterized by hybridization, sequencing and gene synthesis. DNA synthesized in capillaries was capable of being hybridized and signal intensities correlated with microarray data. Sequencing demonstrated that the oligonucleotides were of high quality (up to 44% perfect sequences). Oligonucleotides were combined and used successfully for gene synthesis. This system offers a novel, scalable method to synthesize high quality oligonucleotides for biological applications.
机译:随着合成生物学和基因组科学方面的研究变得越来越广泛,对多样化寡核苷酸群体的需求也在增加。为了限制试剂成本,以微量获得高质量的种群将是有利的。为此,研究了利用光不稳定的3-硝基苯丙氧基羰基(NPPOC)化学物质和紫外发光二极管(UV-LED)在毛细管中合成DNA链的方法。在单个毛细管中制备了多个寡核苷酸,并通过杂交,测序和基因合成进行了表征。在毛细管中合成的DNA能够杂交,并且信号强度与微阵列数据相关。测序证明寡核苷酸是高质量的(高达44%的完美序列)。寡核苷酸被组合并成功用于基因合成。该系统提供了一种新颖的,可扩展的方法来合成用于生物学应用的高质量寡核苷酸。

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