DNA-SEQ of synthetic library for quantifying growth: Minimal media, after 33 generations of competition, replicate #2
The synthetic E. coli library comprising 244,000 variants of a fluorescent reporter was grown in rich MOPS medium for 33 generations upon supplementation with IPTG to induce reporter expression (triplicates). During this time, strains in the library are competing with each other. The variable region of the reporter was PCR amplified from DNA minipreped from the evolved populations. Primers used for generating the sequencing libraries contained handles for sequencing as well as custom sequencing barcodes for multiplexing. Libraries from various evolved populations were pooled and sequenced together (150PE). Resulting reads were demultiplexed through identification of the custom barcodes, which was then trimmed from the sequences along with the priming region. The fastq files contain these trimmed, demultiplexed reads. Reads with no assignable barcode were thrown away. No other data processing was performed.