Mutation Accumulation (MA) of Escherchia coli: WGS of mutL- evolved
Genomic DNA was extracted (GenElute Bacterial Genomic DNA kit, Sigma-Aldrich) from 1 mL of an overnight (~14 hours, shaking at 200 rpm, 37C) LB broth cultures of the final time-point (day 50) clones from each of the 20 evolved replicates from the mutation accumulation (MA) experiment and quantified using the Qubit HS dsDNA assay, Invitrogen. Equal amounts of genomic DNA from each of the 20 evolved lines was pooled into a single tube. A paired-end library was prepared from the pooled genomic DNA using the Illumina Nextera XT DNA library preparation kit per the manufacturers instructions. The library was sequenced on the Illumina MiSeq platform using the 2250bp paired-end V2 reaction chemistry.