Barcode-seq of yeast  Knockout Collection library treated with drug 542
A pool of Yeast Knockout Collection Haploid Mat-a/ (Trans OMIC technologies) was distributed in YPD  medium, either with the drug or vehicle control (DMSO) in 48-well plates. The plates were screened in BioScreen for 3 days and then they were frozen down. Once the cultures with drugs ~IC20s were determined, cultures were harvested by for genomic DNA extraction with LiOAc-SDS protocol. After DNA extraction, a PCR was performed to amplify the barcodes, using for each sample a different tagged forward primer,  for post-sequencing sample identification.  Finally, the PCR product was purified using the Wizard Genomic DNA Purification Kit (SERVER PROMEGA, 2020) and sent for Illumina sequencing (150 bp paired-end reads).