Droplet Tn-Seq combines microfluidics with Tn-Seq for identifying complex single-cell phenotypes.

ABSTRACT: While Tn-Seq is a powerful tool to determine genome-wide bacterial fitness in high-throughput, culturing transposon-mutant libraries in pools can mask community or other complex single-cell phenotypes. Droplet Tn-Seq (dTn-Seq) solves this problem by microfluidics facilitated encapsulation of individual transposon mutants into growth medium-in-oil droplets, thereby enabling isolated growth, free from the influence of the population. Here we describe and validate microfluidic chip design, production, encapsulation, and dTn-Seq sample preparation. We determine that 1-3% of mutants in Streptococcus pneumoniae have a different fitness when grown in isolation and show how dTn-Seq can help identify leads for gene function, including those involved in hyper-competence, processing of alpha-1-acid g

SUBMITTER: Thibault D 

PROVIDER: S-EPMC6914776 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature