Yeast Growth Profiling in Solid and Liquid Media

Solid media plates were cast with 50 ml of Synthetic Complete (SC) medium: 0.14% Yeast Nitrogen Base (YNB, CYN2210, ForMedium), 0.50% ammonium sulfate, 0.077% Complete Supplement Mixture (CSM, DCS0019, ForMedium), 2.0% (w/v) glucose and pH buffered to 5.8 with 1.0% (w/v) succinic acid, and 0.6% (w/v) NaOH. Media was supplemented with 20 g/L of agar. Where indicated, 2.0% glucose was replaced by 2.0% galactose and/or supplemented with salt to 0.85 M NaCl. Three strain layouts were used: 1) all colonies being diploid BY4743 reference strain (Brachmann et al. 1998 (link)) (Figure 1, Figure 2, and Figure 3); 2) colonies corresponding to single yeast gene knockouts of the haploid BY4742 deletion collection (Giaever et al. 2002 (link)), with WT control colonies interleaved in every fourth position and n = 3 replicates of each strain in juxtaposition (Figure 4, A–D); 3) for the confirmation experiment (Figure 4, E and F) the same procedure was employed, but at high number of replicates (n = 24). The reference liquid media experiments (n = 6) (Figure 4, E and F) were performed as previously described (Warringer et al. 2011 (link)). See File S1, section 8, for a complete description of the wet-lab procedures.