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. 2009 Nov 10;4(11):e7775. doi: 10.1371/journal.pone.0007775

Figure 1. Targeting the Mouse Wwox allele.

Figure 1

(A) Insertion of the targeting vector into the Wwox allele introduced a new Bgl II (B) restriction site into Wwox Intron 1. The new Bgl II site leads to a smaller Bgl II restriction fragment when analyzed by Southern blot with the external hybridization probe. Additionally, a Not I restriction site was engineered at the 3′ end of the right arm used to linearize the targeting construct prior to electroporation. (B) Southern blot genotyping of wild-type (wt/wt) mice, heterozygous mice (wt/flox) and homozygous mice (flox/flox). (C) Multiplex-PCR based genotyping using allele specific primers. Primers A and B anneal to Wwox specific sequences in the Wwox upstream regulatory region (primer A) and in Wwox Exon 1 (primer B). Primer C anneals to sequences in the LoxP Cre-recombination in the targeted Wwox allele. Multiplex PCR using primers A, B and C yield DNA products having sizes specific for the wild-type and targeted alleles.