DNA-SEQ of synthetic library for quantifying growth: Regular condition, uninduced reporter expression, before 2nd batch of competition
The synthetic E. coli library comprising 244,000 variants of a fluorescent reporter was first grown in rich MOPS medium without induction of the reporter. A sample was then taken for sequencing to characterize the initial state of the library prior to a second batch of competition experiments. The variable region of the reporter was PCR amplified from minipreped DNA. 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.