Figure 4
Identification of F2 generation homozygous Them2 mutant mice via PAGE-based genotyping protocol.
Mouse #5 (Figure 2D) were bred with wildtype C57/BL6 mice to obtain F1 mice. We further intercrossed mice from F1 to obtain homozygous F2 mutant mice. (A) Identification of F1 offsprings using PAGE-based genotyping approach. Five out of nine F1 offsprings were identified by PAGE analysis as carrying one type of indel mutations in the Them2 locus. (B) Sequencing results from F1 offsprings revealed mice harboring a 3-bp deletion. (C) Identification of F2 offsprings using PAGE-based genotyping approach. F2 mice #3, #4, #5 and #7 harboring heterozygous indel mutations in their Them2 locus were identified by PAGE assay (upper panel). When mixing PCR products from control mice with PCR products from F2 mice, mice #2 and #8 exhibited migration patterns from heteroduplex DNA (marked by asterisks), indicating homozygous indel mutation (bottom panel). M, DNA marker; WT, wildtype; Homo, homozygote; Het, heterozygote.
