SequencingRun2_SK7622_gen10
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 diluted exponentially-growing cultures of the libraries in LB media. Cultures were allowed to grow for ~10 generations at 37C with a 32-fold dilution after ~5 generations of growth. We plasmid prepped and determined the allele frequencies of the generation 10, replica 2 sample using deep sequencing.