title: Genotyping of Scer harboring FCY1 promoter mutations upstream of an mEGFP construct, after FACS
design: A yeast strain harboring a fluorescent tag at the FCY1 locus (keeping only the first 42 bp of the original gene) was transformed with a pool of FCY1 promoters each harboring a single mutations, in three biological replicates. Cells from the resulting libraries of mutants were sorted according into five bins to their fluorescence intensity (dead, weak, medweak, medstrong, strong), twice per library (technical replicates). Genomic DNA was extracted from the collected cells and two rounds of PCR were used to amplify the mutagenized promoter region and flanking bases covering the remaining fragment of FCY1 (42 bp) and the first 85 bp of the mEGFP fluorescent tag.
identifiers.namespace: SUB15610605
identifiers.id: F3F4_Sort