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微芯片上的核酸分离与富集

Nucleic Acid Isolation and Enrichment on a Microchip.

作者信息

Kim Jinho, Hilton John P, Yang Kyung A, Pei Renjun, Stojanovic Milan, Lin Qiao

机构信息

Department of Mechanical Engineering, Columbia University, New York, NY 10027.

Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, New York, NY 10032.

出版信息

Sens Actuators A Phys. 2013 Jun 1;195:183-190. doi: 10.1016/j.sna.2012.07.022.

Abstract

This paper presents a microchip that isolates and enriches target-binding single-stranded DNA (ssDNA) from a randomized DNA mixture using a combination of solid-phase extraction and electrophoresis. Strands of ssDNA in a randomized mixture are captured via specific binding onto target-functionalized microbeads in a microchamber. The strands are further separated from impurities and enriched on-chip via electrophoresis. The microchip consists of two microchambers that are connected by a channel filled with agarose gel. In the isolation chamber, beads functionalized with human immunoglobulin E (IgE) are retained by a weir structure. An integrated heater elevates the temperature in the chamber to elute desired ssDNA from the beads, and electrophoretic transport of the DNA through the gel to the second chamber is accomplished by applying an electric potential difference between the two chambers. Experimental results show that ssDNA expressing binding affinity to IgE was captured and enriched from a sample of ssDNA with random sequences, demonstrating the potential of the microchip to enhance the sensitivity of ssDNA detection methods in dilute and complex biological samples.

摘要

本文介绍了一种微芯片,该微芯片利用固相萃取和电泳相结合的方法,从随机DNA混合物中分离并富集与靶标结合的单链DNA(ssDNA)。随机混合物中的ssDNA链通过特异性结合捕获到微腔中靶标功能化的微珠上。这些链进一步与杂质分离,并通过电泳在芯片上进行富集。该微芯片由两个微腔组成,两个微腔由填充有琼脂糖凝胶的通道相连。在分离腔中,用人免疫球蛋白E(IgE)功能化的微珠被堰结构保留。集成加热器升高腔室内的温度,以从微珠上洗脱所需的ssDNA,通过在两个腔室之间施加电势差,使DNA通过凝胶电泳传输到第二个腔室。实验结果表明,从具有随机序列的ssDNA样本中捕获并富集了对IgE具有结合亲和力的ssDNA,证明了该微芯片在提高稀释和复杂生物样本中ssDNA检测方法灵敏度方面的潜力。

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Nucleic Acid Isolation and Enrichment on a Microchip.微芯片上的核酸分离与富集
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