Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
Methods Enzymol. 2020;643:317-343. doi: 10.1016/bs.mie.2020.06.002. Epub 2020 Jul 24.
Water-in-oil droplets, made and handled in microfluidic devices, provide a new experimental format, in which ultrahigh throughput experiments can be conducted faster and with minimal reagent consumption. An increasing number of studies have emerged that applied this approach to directed evolution and metagenomic screening of enzyme catalysts. Here, we review the considerations necessary to implement robust workflows, based on choices of device design, detection modes, emulsion formulations and substrates, and scope out which enzyme classes have become amenable to droplet screening.
油包水液滴在微流控装置中生成和处理,提供了一种新的实验形式,可以更快地进行超高通量实验,并且试剂消耗最小。越来越多的研究采用这种方法进行酶催化剂的定向进化和宏基因组筛选。在这里,我们根据设备设计、检测模式、乳剂配方和底物的选择,回顾实施稳健工作流程所需的注意事项,并概述哪些酶类已适用于液滴筛选。