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Fig. 3 | BMC Genomics

Fig. 3

From: One-step generation of a targeted knock-in calf using the CRISPR-Cas9 system in bovine zygotes

Fig. 3

Identification of allelic sequence at the H11 target site. The coverage depth was calculated for the mapped alignment of Illumina NovaSeq whole genome sequencing reads to the expected knock-in, the Sanger sequenced 26 bp knock-in allele, and the pUC19 donor plasmid backbone. Reads were then used to identify the junction sites between the insertions. a coverage depth of reads aligned to the complete 5.1 kb SRY-GFP template b coverage depth of reads aligned to the 26 bp insertion, c coverage depth of reads aligned to the 2.7 kb pUC19 donor plasmid backbone (orange), and d the 38 kb and 18 kb complex insertions, and e gRNA target site and Cas9 cut site (yellow) at the H11 locus on chromosome 17, and schematic representation of the 38 kb and 18 kb complex allele insertion junctions (1–10). LHA = left homology arm; SRY = sex-determining region Y; GFP = green fluorescent protein; RHA = right homology arm; pUC = pUC19 donor plasmid backbone; CHR17 = genomic region outside homology arms on chromosome 17; KI = complete 5.1 kb SRY-GFP template; tLHA = truncated left homology arm; PAM = protospacer adjacent motif

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