Comparison of methods to generate a pooled Enterococcus faecalis Tn library from arrayed library plates

The goal of this study was to determine how the method of pooling (combining mutants) for bacteral Tn libraries affected mutant distribution. Previously we generated a pooled version of a~6,800 Tn mutant library in Enterococcus faecalis OG1RF by plating and scraping individual mutants. Here, we combined liquid cultures grown in deep well plates. DNA from the new pooled library was extracted, and TnSeq was used to compare mutant distribution between the old (plate scraping) and new (liquid pooling) Tn library formats. This is important because it will guide best practices for handling large collections of bacterial mutants. Overall design: TnSeq using purified genomic DNA from bacterial samples