Whole genome sequencing of E. coli evolved to thermal stress
To sequence the evolved populations, we revived population from ~10 l of frozen glycerol stock in 10ml of DM media supplemented with 1000mg/L of glucose. The culture tubes were incubated at 19.0C with 120 RPM. We extracted from 65 bacterial populations using the Promega Wizard Genomic DNA Purification kit. DNA concentrations were measured with Qubit dsDNA HS Assay kits. We prepared our DNA sequencing libraries with the Illumina Nextera DNA Flex Library Preparation kit. The libraries were multiplexed and sequenced using the Illumina NovaSeq on an S4 flow cell to generate 100bp paired-end reads at UC Irvines Genomics High-Throughput Facility