Fig. 3

Characterization of NHEJ-mediated donor knock-in at Alb stop codon. a Schematic of forward and reverse integrations of the BDDF8 donor or plasmid backbone. Eight pairs of primers were designed to amplify the junctions (F8a, F8b, F8c, F8d, BB1, BB2, BB3, and BB4). The red arrow indicates the sgAlb target site. b Successful amplification of the eight junctions using designed primers. Shown is a representative result using live gDNA from one edited mouse. The identity of these PCR products was confirmed by Illumina sequencing (Additional file 1: Fig. S6). c A representative diagram of ddPCR analysis of the copy number of NHEJ-mediated knock-in. One hundred nanograms of gDNA was used in each reaction. To count the total number of haploid genomes interrogated, we used a probe that targets the Actb gene. d Quantitation of copy numbers for the eight junctions are shown. F8fwd, insertion of BDDF8 in the forward orientation; F8rev, insertion of BDDF8 in the reverse orientation; BBfwd, insertion of plasmid backbone in the forward orientation; BBrev, insertion of plasmid backbone in the reverse orientation