Difco yeast nitrogen base without amino acids and ammonium sulfate (YNB), Bacto peptone, Bacto yeast extract, Luria Broth (LB), LB agar, dextrose, and galactose were obtained from BD (Becton, Dickinson and Company). PBS tablets were obtained from VWR. BSA was obtained from Cell Signaling Technologies. kanamycin monosulfate and ampicillin were obtained from Sigma Chemicals. G418 (sc-29065A) used for yeast integration selections was obtained from Santa Cruz Biotech. Amino acid dropout media was obtained from Takara Bio (product #630400–#630431). 5-FOA powder and Frozen-EZ transformation kit for yeast were obtained from Zymo Research.
E. coli strains were selected on LB agar plates with 50 mg/L kanamycin, 50 mg/L ampicillin, and grown in LB liquid media with the appropriate antibiotic. Yeast 10x drop out (DO) supplement was prepared from Takara as a synthetic complete supplement with desired dropout component omitted. S. cerevisiae strains were selected on YNB-DO (0.17% yeast nitrogen base, 0.5% ammonium sulfate, 2% dextrose, and 1x DO) agar or on YPAD (1% yeast extract, 2% peptone, 80 mg/L adenine hemisulfate, and 2% dextrose) agar. For selections, plates were supplemented with 200 mg/L G418 or 400 mg/L 5-FOA. Yeast were grown in selective YNB-DO media or in YPAD media.
E. coli strains were selected on LB agar plates with 50 mg/L kanamycin, 50 mg/L ampicillin, and grown in LB liquid media with the appropriate antibiotic. Yeast 10x drop out (DO) supplement was prepared from Takara as a synthetic complete supplement with desired dropout component omitted. S. cerevisiae strains were selected on YNB-DO (0.17% yeast nitrogen base, 0.5% ammonium sulfate, 2% dextrose, and 1x DO) agar or on YPAD (1% yeast extract, 2% peptone, 80 mg/L adenine hemisulfate, and 2% dextrose) agar. For selections, plates were supplemented with 200 mg/L G418 or 400 mg/L 5-FOA. Yeast were grown in selective YNB-DO media or in YPAD media.