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. 2006 Jan;172(1):709–711. doi: 10.1534/genetics.105.047662

Figure 1.

Figure 1.

Sources of variability in SGA. (A) MATα cells of strain Y2454 from an overnight culture grown in YPD were spread onto an SC–his plate and incubated for three days at 30°. A small region is shown. Approximately 1000 colonies of varying sizes were evident on the entire plate. (B) The same region after replica plating to a plate containing 2 mm 3-aminotriazole, which inhibits the growth of the small colonies. Arrows point to the same colonies on both plates. (C) Haploid revertants were recovered and tested for gene conversion at the his3Δ1 locus using primers flanking the gene. Lane M is molecular weight marker, lane B is genomic DNA from BY4741 (his3Δ1) as template, lane P is plasmid pRS303 (HIS3) DNA as template, and 1–15 are DNA from the 15 revertants. All of the revertants were MATa except revertant 3, which was MATα. (D) False negative colonies were recovered from SGA after using PMFA1–HIS3 in the query strain and tested for gene conversion at the his3Δ1 locus, using primers flanking the gene. Lane M is molecular weight marker, lane B is genomic DNA from BY4741 (his3Δ1) as template, lane P is plasmid pRS303 (HIS3) DNA as template, and 1–15 are DNA from the 15 revertants. All of the revertants were MATa except 1–4, which were MATa/MATα .