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用于大规模平行原位合成寡核苷酸的微流控反应器阵列装置

Microfluidic Reactor Array Device for Massively Parallel In-situ Synthesis of Oligonucleotides.

作者信息

Srivannavit Onnop, Gulari Mayurachat, Hua Zhishan, Gao Xiaolian, Zhou Xiaochuan, Hong Ailing, Zhou Tiecheng, Gulari Erdogan

机构信息

Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.

出版信息

Sens Actuators B Chem. 2009 Jul;140(2):473-481. doi: 10.1016/j.snb.2009.04.071.

Abstract

We have designed and fabricated a microfluidic reactor array device for massively parallel in-situ synthesis of oligonucleotides (oDNA). The device is made of glass anodically bonded to silicon consisting of three level features: microreactors, microchannels and through inlet/outlet holes. Main challenges in the design of this device include preventing diffusion of photogenerated reagents upon activation and achieving uniform reagent flow through thousands of parallel reactors. The device embodies a simple and effective dynamic isolation mechanism which prevents the intermixing of active reagents between discrete microreactors. Depending on the design parameters, it is possible to achieve uniform flow and synthesis reaction in all of the reactors by proper design of the microreactors and the microchannels. We demonstrated the use of this device on a solution-based, light-directed parallel in-situ oDNA synthesis. We were able to synthesize long oDNA, up to 120 mers at stepwise yield of 98 %. The quality of our microfluidic oDNA microarray including sensitivity, signal noise, specificity, spot variation and accuracy was characterized. Our microfluidic reactor array devices show a great potential for genomics and proteomics researches.

摘要

我们设计并制造了一种用于大规模并行原位合成寡核苷酸(oDNA)的微流控反应器阵列装置。该装置由阳极键合到硅上的玻璃制成,具有三个层次的特征:微反应器、微通道和贯通的进出口孔。设计该装置的主要挑战包括在激活时防止光生试剂扩散,以及实现试剂在数千个平行反应器中的均匀流动。该装置体现了一种简单有效的动态隔离机制,可防止离散微反应器之间的活性试剂相互混合。根据设计参数,通过对微反应器和微通道进行适当设计,有可能在所有反应器中实现均匀流动和合成反应。我们展示了该装置在基于溶液的光导向并行原位oDNA合成中的应用。我们能够以98%的逐步产率合成长达120个碱基的长oDNA。对我们的微流控oDNA微阵列的质量进行了表征,包括灵敏度、信号噪声、特异性、斑点变化和准确性。我们的微流控反应器阵列装置在基因组学和蛋白质组学研究中显示出巨大潜力。

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