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Fig. 4 | BMC Biology

Fig. 4

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

Fig. 4

Screening by Sanger sequencing of animals for the generation of the GckrP446L point mutation with (a) oligonucleotides (F0 individual ssO-GckrP446L-54) or (b) lssDNA donors (F0 individuals lss-GckrP446L-11 and lss-GckrP446L-10 and F1 individual lss-GckrP446L-11.1f). The figure shows Sanger sequencing chromatograms of an amplicon generated with primers anchored external to the intended site of donor sequence integration as detailed in Additional file 15: Figure S14. a ssODN donors only yielded introduction of the intended silent mutations, while (b) lssDNA yielded the desired mutation in some individuals (F0 11 transmitting to 11.f) and only the silent mutations in others (F0 10). Note that founders appeared homozygous (ssO-GckrP446L-54, lss-GckrP446L-11 and lss-GckrP446L-10) when analyzed by Sanger sequencing, but also could contain deletion alleles in trans, as suggested by copy counting (lss-GckrP446L-11 in Table 4). A summary of the microinjection session outcomes is detailed in Table 3, and raw sequencing data are provided in Additional file 16

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