Droplet-Based Microfluidic Systems for High-Throughput Single DNA Molecule Isothermal Amplification and Analysis
Droplet-Based Microfluidic Systems for High-Throughput Single DNA Molecule Isothermal Amplification and Analysis
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DOI：
10.1021/ac900403z
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发表时间：
2009-06-15
影响因子：
7.4
通讯作者：
Ryckelynck, Michael
中科院分区：
文献类型：
--
作者：
Mazutis, Linas;Araghi, Ali Fallah;Ryckelynck, Michael
我们开发了一种基于微流控系统的高通量等温扩增单DNA分子的方法。使用插层荧光染料分析液滴中的DNA扩增，允许基于液滴中DNA分子的泊松分布对模板DNA进行快速准确的“数字”量化。每个2 pL液滴中克隆扩增的DNA与含有体外翻译系统的15 pL液滴融合后，通过测量编码蛋白的酶活性进一步分析。
We have developed a method for high-throughput isothermal amplification of single DNA molecules in a droplet-based microfluidic system. DNA amplification in droplets was analyzed using an intercalating fluorochrome, allowing fast and accurate "digital" quantification of the template DNA based on the Poisson distribution of DNA molecules in droplets. The clonal amplified DNA in each 2 pL droplet was further analyzed by measuring the enzymatic activity of the encoded proteins after fusion with a 15 pL droplet containing an in vitro translation system.