Fig. 1.

Strategy used to generate diploid segregant pools. Diploid evolved clones isolated from Generation 2000 were sporulated and mated en masse to barcoded MATα version of the ancestor. Each unique barcode represents a single mating event. The resulting diploids have genotypes intermediate to their ancestor and evolved parents and carry a barcode. Mutations homozygous in the evolved parent (in orange) end up in heterozygosity in all segregants. Supplementary Fig. 1 shows the barcoded locus.