Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR
Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR
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用于高度平行且高效的单分子乳液 PCR 的琼脂糖液滴微流控
DOI：
10.1039/c0lc00145g
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Yang, Chaoyong James
中科院分区：
文献类型：
--
作者：
Leng, Xuefei;Zhang, Wenhua;Yang, Chaoyong James
建立了一种高效、并行的单分子乳状液聚合酶链式反应的琼脂糖滴法。该方法利用琼脂糖热响应性独特的溶胶-凝胶开关特性进行高效的DNA扩增和扩增捕获。通过微流控芯片产生的均匀的琼脂糖液滴可作为单拷贝DNA分子扩增的坚固和惰性纳米级聚合酶链式反应。聚合酶链式反应后，琼脂糖滴被凝胶形成琼脂糖珠，捕获每个反应器中的所有扩增，以保持每个液滴的单克隆性。这种方法不需要对标记的微球进行包埋，可以高通量地产生均匀的液滴，并且能够实现高的聚合酶链式反应效率，使其成为许多单拷贝遗传研究的一个有前途的平台。
An agarose droplet method was developed for highly parallel and efficient single molecule emulsion PCR. The method capitalizes on the unique thermoresponsive sol-gel switching property of agarose for highly efficient DNA amplification and amplicon trapping. Uniform agarose solution droplets generated via a microfluidic chip serve as robust and inert nanolitre PCR reactors for single copy DNA molecule amplification. After PCR, agarose droplets are gelated to form agarose beads, trapping all amplicons in each reactor to maintain the monoclonality of each droplet. This method does not require cocapsulation of primer labeled microbeads, allows high throughput generation of uniform droplets and enables high PCR efficiency, making it a promising platform for many single copy genetic studies.