DMS of NEB 5-alpha F'Iq: Single-codon substitutions in NDM-1, sequencing run 2, generation 10
A library of single-codon substitutions in NDM-1 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 NDM-1 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 2 sample using deep sequencing. Data for codons 1-271 was used from these files. This was used as Replica 2 for regions 1 (codons 1-90), 2 (codons 91-180), and 3 (codons 181-271).