Ultrahigh-Throughput Screening of Metagenomic Libraries Using Droplet Microfluidics

Abstract

Droplet microfluidics enables the ultrahigh-throughput screening of the natural or man-made genetic diversity for industrial enzymes, with reduced reagent consumption and lower costs than conventional robotic alternatives. Here we describe an example of metagenomic screening for nucleoside 2'-deoxyribosyl transferases using FACS as a more widespread and accessible alternative than microfluidic on-chip sorters. This protocol can be easily adapted to directed evolution libraries by replacing the library construction steps and to other enzyme activities, e.g., oxidases, by replacing the proposed coupled assay.

Keywords: Enzyme screening; Functional metagenomics; Microfluidics; Nucleoside 2′-deoxyribosyl transferases.