SequencingRun2_SK7622_gen0
A library of single-codon substitutions in the rnc gene was constructed using inverse PCR. The library was transformed into NEB 5-alpha cells. Plasmids were prepped from NEB5alpha colonies that grew overnight on bioassay plates. That plasmid library was transformed into SK7622 cells and colonies that grew overnight diluted on bioassay plates were collected.  We plasmid prepped exponentially-growing cultures of the library mixed and determined the allele frequencies at the start of the growth experiment as our generation 0, replica 2 sample using deep sequencing.