Additional file 5: of Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants

Figure S4. Analysis of the Ikzf2 project. PCR amplification of the genomic region of interest from (a, b) F0 animals and (f, g) Ikzf2-2’s offspring with (a, f) Ikzf2-F3 and Ikzf2-3R2 primers (1594-bp amplicon) and (b, g) LoxPF and LoxPR primers (906-bp amplicon) from biopsies. (a, b, f, g) Animals’ IDs are shown. + is positive control amplified from an unrelated (a) WT, (b) plasmid template. Sequencing of PCR amplicon from (c) the founder Ikzf2-2, (h) Ikzf2-2.1f and (i) Ikzf2-2.1 h with Ikzf2-F3 and Ikzf2-3R2 primers. LoxP sequences are highlighted in blue. (d) ID and outcome of PCR analysis of the region of interest and the conclusion for each F0 individual. (e) ID, outcome of sequencing and copy counting of the region of interest as well as the conclusion for each individual of the first litter obtained by mating Ikzf2-2 with a WT mouse. *Animal mated; **deletion not picked up by Ikzf2 PCR, likely encompassing at least one primer sequence; ***allele detailed in Additional file 14: Figure S13. Evidence of deletion is highlighted in blue. L1 = 1 kb DNA molecular weight ladder (thick band is 3 kb). Sequencing data showing a correct conditional allele are shown in Additional file 3: Figure S2d. Sequencing data showing the presence of a deletion allele in founders Ikzf2-4 and Ikzf2-8 are shown in Additional file 3: Figure S2b and c. (PNG 1031 kb)