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. 2023 Jul 20;13(14):e4724. doi: 10.21769/BioProtoc.4724

Figure 4. PCR validation of PD-L1 knockout founder pups.

Figure 4.

Target gene was amplified and subsequently digested with HindIII restriction enzyme. Numbers 1–19 represent the different founder pups. The left side shows DNA ladder with depicted sizes. 400 bp bands show wildtype alleles, while 200 bp bands represent cut alleles that integrated the ssDNA donor template with HindIII restriction site. Subsequent digestion with HindIII restriction enzyme after PCR cuts the alleles harboring the de novo integrated HindIII 200 bp fragments. Heterozygous pups show two bands; homozygous pups show one band. Only pup number 10 has both mutated alleles.