Schematic design of the SSM Schematic design of the SSM. (A) The genetic crosses required for generating multiple independent m/m and sibling +/+ isolates, which are combined to produce the mutant DNA pool and the nonmutant sibling comparator DNA pool, respectively (also see Figure S3). Schematic design of the SSM. (A) The genetic crosses required for generating multiple independent m/m and sibling +/+ isolates, which are combined to produce the mutant DNA pool and the nonmutant sibling comparator DNA pool, respectively (also see Figure S3). (B) Simplified representations of sequenced chromosomes from the mutant pool (pink) and nonmutant sibling comparator pool (blue) are shown. Yellow lines indicate variants detected by WGS, and asterisks indicate variants that are homozygous in the mutant pool. Note that the depiction underestimates the true number of variants per chromosome. Dashed lines and purple asterisks indicate variants that are present in both the mutant pool and the nonmutant sibling comparator pool, which can be eliminated (subtracted) as candidate causal mutations. Note that subtraction will remove most or all variants on unlinked chromosomes, whereas homozygous variants very close to the causal mutation (red) may not be subtracted (black asterisks). (C) Venn diagram of subtracted variants (purple) along with the relatively small proportion of remaining candidate variants (pink) after application of the SSM. Braveen B. Joseph et al. G3 2017;8:669-678 ©2018 by Genetics Society of America