E. coli barcoded deletion library screen for chemotherapeutic antimetabolite gemcitabine

A genetic screen was performed using barcoded E. coli genome-wide knockout library and sub-inhibitory concentration of antimetabolite drug gemcitabine. This knockout library contains 3,680 mutants, each having a unique DNA barcode, GFP tag and chloramphenicol resistance gene in the knockout locus. The library was generated from the parent strain BW38028. Pooled library was grown in the presence and absence of gemcitabine in M9 miminal media as biological triplicates, and the experiment was stopped when the cultured reached OD600 value of 0.6. Genomic DNA was extracted and the barcodes were amplified and sequenced by Illumina Next Generation Sequencing.