title: Genotyping of Scer harboring FCY1 promoter mutations after bulk competition, using an extended sequencing region
design: S. cerevisiae yeasts transformed with a pool of FCY1 promoters each harboring a single mutations were grown in SC media with or without 5-FC [12 uM] for 20 doublings, with intermediate dilution steps to maintain exponential growth (T0: start; T1: first dilution; T2: second dilution; T3: end). Each pool of mutants was generated in four biological replicates, each used to inoculate two technical replicates. From all collected cell pellets, genomic DNA was extracted and two rounds of PCR were used to amplify the 154 bp mutagenized region and flanking bases covering the first 43 codons of the FCY1 coding sequence.
identifiers.namespace: SUB15609779
identifiers.id: F3F4_300