WGS of E. coli K-12 pooled transposomes
A previously published toehold-mediated reporter plasmid was transformed into EZ-Tn5 generated E. coli transposomes. Cells were sorted based on fluorescence levels using a FACSAria Illu cell sorter. After isolation and low-throughput validation, overnight cultures were started for each of the 64 library isolates and genomic DNA (gDNA) was extracted using a QIAamp UCP DNA Micro Kit (QIAGEN) and following manufacturers instructions. gDNA was extracted from wild type E. coli MG1655 cells as a control. Genomic DNA was submitted to the GSAF core facility (UT Austin) for library prep and sequencing. DNA libraries were prepared using standard Illumina kits and were run using a MiSeq sequencer (Illumina) in a 2x250 paired-end scheme.