GSM4039954: NBT_S288CdU_YPD_R15; Saccharomyces cerevisiae; OTHER
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 (except for scaffold library, where bins were approximarely proportional to the expression variance within the bin), 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 SC-Ura (supplemented with 1% Gal for Gal sort) and grown for 2-3 days, shaking at 30C. 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. The scaffold library was sequenced with 300 cycle kits in two reads of 190 and 110 bases. The second replicate of the Native80 library was sequenced with a 300 cycle kit.