A Simultaneous Space-sampling Method for DNA Fraction Collection Using a Comb Structure in Microfluidic Devices
A Simultaneous Space-sampling Method for DNA Fraction Collection Using a Comb Structure in Microfluidic Devices
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微流控装置中使用梳状结构的 DNA 组分同步空间采样方法
DOI：
10.1002/elps.201100362
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发表时间：
2011
期刊：
影响因子：
2.9
通讯作者：
Zheyu Li
中科院分区：
文献类型：
--
作者：
Shibata;N.;Hayashi;T.;Fukumura;R.;Fujii;J.;Kudome-Takamatsu;T.;Nishimura;O.;Sano;S.;Son;F.;Suzuki;N.;Araki;R.;Abe;M.;Agata;K.;Zheyu Li
从复杂的混合物中收集选定的组分在生物医学研究、环境分析和生物技术中起着关键作用。在这里，我们介绍了一种新的电泳芯片设备的基础上的信号处理定理，允许同时空间采样的ssDNA靶片段的分馏。使用了10个平行的提取通道，覆盖1.5 mm长的采样范围，以便于捕获快速移动的碎片。此外，空间采样提取使得有可能获得纯收集，甚至从部分重叠的片段，已经在短的电泳运行后没有充分分离。同时收集180、181和182个碱基的片段，然后对回收的DNA进行PCR扩增并通过CE分析进行评估。181个碱基的靶显示在10分钟内在70 mm长的分离长度中被分离，而在我们之前的研究中，300 mm长的分离通道需要>50分钟。该方法提供了时间和空间的有效结合，这是对传统的芯片上的馏分收集的概念的突破。
Fraction collection of selected components from a complex mixture plays a critical role in biomedical research, environmental analysis, and biotechnology. Here, we introduce a novel electrophoretic chip device based on a signal processing theorem that allows simultaneous space sampling for fractionation of ssDNA target fragments. Ten parallel extraction channels, which covered 1.5‐mm‐long sampling ranges, were used to facilitate the capturing of fast‐moving fragments. Furthermore, the space sampling extraction made it possible to acquire pure collection, even from partly overlapping fragments that had been insufficiently separated after a short electrophoretic run. Fragments of 180, 181, and 182 bases were simultaneously collected, and then the recovered DNA was PCR amplified and assessed by CE analysis. The 181‐base target was shown to be isolated in a 70‐mm‐long separation length within 10 min, in contrast to the >50 min required for the 300‐mm‐long separation channel in our previous study. This method provides effective combination of time and space, which is a breakthrough in the traditional concept of fraction collection on a chip.