E. coli barcoded deletion library screen for antimetabolite chemotherapies 5-FU and FUDR resistance in nutrient-poor & rich media

An E. coli barcoded deletion library was used for a genetic screen of resistance to antimetabolite chemotherapies 5-FU and FUDR in nutrient-poor or nutrient-rich media. The parent strain of the library is BW38028. This library consists of 3,680 individual knockout strains with their own unique 20 bp sequence serving as an identification barcode. The screen using the pooled library was performed with biological duplicates of each drug/media combination and the screen was stopped when E. coli populations were in exponential growth phase. The barcoded regions were amplified in each screen sample as well as the initial inoculum population, and this barcode amplicon product was used for Illumina NGS.