Amplicon sequencing of M. smegmatis after oligonucleotide recombination with a collection of oligonucleotides containing 4 mismatches
Electrocompetent M. smegmatis that had been expressing Che9c gene 61 recombinase is electroporated with a collection of oligonucleotides that introduce mismatches that should generate a rifampicin resistance mutation in the gene rpoB and other mismatches, allowed to recover in liquid media, then rifampicin is added to liquid media and the culture allowed to grow for several days. Genomic DNA is harvested and the region around the oligonucleotide binding site is PCR'ed for amplicon sequencing.