Figure 2.

Genotyping of blastocysts. (a) Schematic representation of the used semi-nested design for genotyping. Introns are shown in blue, exons are shown in pink, and the CRISPR target site is shown in yellow. DNA isolated from blastocysts was subjected to PCR with primers (shown as arrows) adjacent to the CRISPR target site. The resulting amplicon was used in a second round of PCR with the same reverse and a nested inner forward primer to generate a 417 bp length product from the WT allele and a 370 bp product from the D47 allele. (b) Representative image of amplicons from semi-nested PCR visualized on agarose gel. The upper and lower band correspond to WT and D47 allele, respectively. Full-length agarose gel is included in the Supplementary Materials (Figure S7).