FIG 3 .

Genotypic analyses and sequencing of C. lusitaniae diploid transformants. (A) PCR assays for identifying ade2 or wild-type (WT) genotypes, with arrows indicating the relative positions of the primers used (see Data Set S1 for sequences). (B) Results of genotypic analyses for red and white diploid colonies, indicating that most red colonies exhibit replacement of ADE2 with the SAT1 gene. However, two red colonies (colonies 1 and 8) had positive SAT1 junction checks but still contained the ADE2 ORF, indicating that one allele was repaired via homologous recombination and the other was repaired via NHEJ. P, parent strain (CAY5019); N, negative control (no DNA). (C) Sequencing of the ADE2 ORF in colonies 1 and 8 shows that mutagenesis via NHEJ resulted in a 1- or 2-nucleotide deletion within the protospacer preceding the PAM sequence (red box).