A rotary microsystem for simple, rapid and automatic RNA purification
A rotary microsystem for simple, rapid and automatic RNA purification
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DOI：
10.1039/c2lc40487g
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发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Seo, Tae Seok
中科院分区：
文献类型：
--
作者：
Park, Byung Hyun;Jung, Jae Hwan;Seo, Tae Seok
在这项工作中，我们展示了一种新的旋转微系统，用于简单，快速和自动化的流感病毒RNA纯化。该微装置由用于RNA捕获的硅溶胶-凝胶基质和用于RNA样品（RS）、洗涤溶液（RW）和洗脱缓冲液（RE）的三个储器组成，所述储器与不同尺寸的微流体通道（RS为120 mm，RW为40 mm，RE为20 mm）连接。利用PDMS的疏水性和微通道的窄缝作为被动的毛细管微阀，通过离心力控制溶液的加载。将5 μ L甲型H1N1流感病毒的裂解物样品、洗涤溶液和洗脱缓冲液注入每个指定的容器中，并将病毒样品、洗涤溶液和洗脱缓冲液以1600、2000和2500 RPM顺序加载到溶胶-凝胶室中，使得病毒RNA能够被捕获在溶胶-凝胶固相中，纯化，RNA捕获率为80%左右，通过逆转录酶PCR成功地从回收的纯化H1N1病毒RNA中扩增出H1和M基因。这种新型的旋转样品制备系统消除了任何复杂的硬件和人为干预，并且以高速和高保真度进行RNA提取。
In this work, we demonstrate a novel rotary microsystem for simple, rapid and automatic influenza viral RNA purification. The microdevice consists of a silica sol-gel matrix for RNA capture, and three reservoirs for a RNA sample (RS), a washing solution (RW) and an elution buffer (RE) that were connected with different dimensional microfluidic channels (120 mm for RS, 40 mm for RW, and 20 mm for RE). The hydrophobic property of PDMS and the narrow microchannel served as a passive capillary microvalve, and the loading of the solutions were controlled by centrifugal force. 5 mu L of a lysate sample of influenza A H1N1 virus, a washing solution and an elution buffer were injected in each designated reservoir, and the virus sample, the washing solution, and the elution buffer were sequentially loaded into the sol-gel chamber at 1600, 2000, and 2500 RPM, enabling the viral RNA to be captured in the sol- gel solid phase, purified, and eluted in 5 min. The RNA capture yield was measured as similar to 80%, and the H1 and M gene were successfully amplified from the recovered purified H1N1 viral RNA by reverse-transcriptase PCR. Such a novel rotary sample preparation system eliminates any complicated hardware and human intervention, and performs the RNA extraction with high speed and high fidelity.