Null alleles of Ak040954 and Gm44386 genes generated by the i-GONAD method. (A,B) Genotyping results of Ak040954 (A) and Gm44386 (B) for the F0 generation using primer pairs shown in C and D. The mice #5 to #9 in A and mice #2, #5, #6, and #9 in B showed large deletions. The star in B indicates a faint, nonspecific band. Note that WT or floxed alleles are too large to be detected in current PCR conditions. (C,D) Gene structures and positions of gRNA and genotyping primers for Ak040954 (C) and Gm44386 (D) genes. (E,F) Sanger sequencing results of founder #9 and #6 as shown in A and B, respectively. For all panels: (WT) wild-type, (N) negative control (water), green ID highlights founders with floxed alleles, (arrow) DNA marker lane, mice with single-side loxP integration are indicated by a bar over the ID, (Δ) deletions, (▿) large insertions.
