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Fig. 5 | CABI Agriculture and Bioscience

Fig. 5

From: Technical considerations towards commercialization of porcine respiratory and reproductive syndrome (PRRS) virus resistant pigs

Fig. 5

Process and scaled screening steps of first-generation pigs (E0). Screening steps of primary generation of pigs farrowed from surrogate is described. DNA from tail samples of all live born E0 pigs is prepared and used for short (Illumina) and long (Oxford Nanopore) amplicon sequencing. A reduced number of pigs containing desired modified CD163 allele are then subsequently sequenced using hybridization capture to verify presence of on-target modified CD163 allele and identify gene editing reagent-mediated off-target INDELs. Selected E0 pigs are then crossed to wild-type, line identical, high genetic merit mates. A description of the DNA sequencing methods used in this process are outlined below

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