Barcode amplicon sequencing of  38 E. coli barcoded deletion strains: growth rich media day1; Biological Duplicate 05
38 E. coli barcoded deletion strains were individually picked from a glycerol stock stored at -80 _C and grown overnight in LB supplemented with 20 _g/mL chloramphenicol. The individual cultures were then diluted to a density of 109 cells/mL. Equal volumes were combined and inoculated into LB at a final density of 107 cells/mL. The assemblies were grown in a 96 deep -well plate containing 1 mL of LB per well at 37 C and were shaken (orbital) at 200 rpm. After 12 hours, the assemblies were thoroughly mixed and 5 _L were transferred to 995 _L of fresh LB. The remaining culture was stored at -80 C for later DNA extraction. This 12-hour cycle was repeated for 14 days to track changes in the assembly composition over time. For days 1, 3, 7 and 14, a region of ~350 bp around the barcode locus was amplified with custom forward and reverse primers using 2x KAPA HiFi HotStart ReadyMix (Kapa Biosystems, Cat#KK2602) to produce the amplicon libraries for each sample. Sequencing was performed using MiniSeq High Output Reagent Kit, 150-cycles (Illumina, Cat# FC-420-1002) on Illumina MiniSeq device.