Whole genome sequencing of TLO mutant: bulk cell
Colonies containing the desired mutation had genomic DNA (gDNA) extracted using the Zymo Quick-DNA Miniprep kit (Zymo Research, Irvine, CA, USA). DNA concentration was established using the Qubit 3 Fluorometer with the dsDNA Broad Range Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Finally, gDNA libraries were built using the NEBNext Ultra II FS DNA library prep kit for Illumina (New England Biolabs, Ipswich, MA, USA) prior to whole genome sequencing (WGS) or submitted as gDNA for preparation by The   Ohio State University Applied Microbiology Services Laboratory (AMSL)) or the University of Wisconsin Biotechnology Center DNA Sequencing Facility (UWBC).