Fig. 2

Duplication of the Chr3-1 and Chr3-2 region. The Chr3-1 and Chr3-2 regions of Chromosome 3 were chosen for the initial experiments. Both duplicating modules were prepared so as to be marked with CgHIS3 and CEN4+CgHIS3, respectively. After transformation, two chromosomes of 158 kb (Fig. 2a) and 160 kb (Fig. 2b) were expected to be generated from Chr3-1 and Chr3-2, respectively. The left and right panel of Fig. 2a and Fig. 2b are PFGE along with the corresponding Southern blot analysis of wild type SJY30; 2 and 4 transformants selected from the conventional PCDup experiment for the Chr3-1 and for the Chr3-2 region, respectively and 8 transformants randomly selected from CRISPR-PCDup for both the Chr3-1 and Chr3-2 regions. Right panel of Fig. 2a and Fig. 2b shows the results of Southern blot analysis for detecting the 317 kb Chromosome 3 and newly duplicated 158 kb and 160 kb chromosomes, respectively