DMS of NEB 5-alpha F'Iq: Single-codon substitutions in AadB, sequencing run 5, generation 10
A library of single-codon substitutions in AadB was constructed using inverse PCR. The library was transformed into NEB 5-alpha F'Iq cells. We diluted uninduced, exponentially-growing cultures of the libraries mixed with wildtype AadB into LB media containing IPTG. 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 1 sample using deep sequencing. Data for codons 1-178 was used from these files. This was used as Replica 1 for regions 1 (codons 1-89) and 2 (codons 90-178).