Fig. 4

PCR-screen of mutants and wild type Neonectria ditissima. Primer Set 1 (Table 2) were used to screen for mutants. A size increase from 3548 bp to 4459 bp would be indicative of a gene disruption. Plasmid DNA, Wild Type genomic DNA, and water were included as controls. A 1-Kb ladder (Thermo Fisher) was included in the first and last lane. A Though the mutants grew on the antibiotic hygromycin, all the mutants screened by PCR did not show an increase in the size of the gene. Thus, all the mutants generated were considered ectopic integrants. B The mutants grew on the antibiotic hygromycin and showed an increase on the size of the gene, typical of a gene disruption. These mutants were generated by the inclusion of 100 mM hydroxyurea in the protoplast buffer