GSM4971235: N80_SC-Ura_Y8203_R6; Saccharomyces cerevisiae; OTHER
Plasmids were isolated using Qiagen spin miniprep kits, as adapted for yeast according to the manufacturer's website. Cells were sorted by the log ratio of RFP to YFP signal (using mCherry and GFP absorption/emission) on a Beckman-Coulter MoFlo Astrios, using 18 uniform bins, done in three batches of six bins each. We note that the FACS configuration varied between experiments (e.g. different laser intensities), resulting in different baseline expression values. Post sort, cells were spun down and resuspended in SD-Ura and grown for 2-3 days, shaking at 30C until cultures reached saturation. The plasmids were then isolated, the promoter region amplified, Nextera adaptors and multiplexing indices added, and the resulting libraries sequenced paired-end, 76 bp each, using 150 cycle kits so that complete coverage of the promoter could be achieved, including overlap in the center. OTHER: amplicon sequencing