Southern Blot analysis of transgenic animals.

<p>Genomic DNA isolated from various transgenic founder lines (57, 457) and wild type (WT) littermates were digested with PstI, separated by agarose gel electrophoresis and (A) autoradiographed after hybridization to a radioactively labeled amelotin cDNA probe isolated from the EcoRV-PstI 528 bp region of the transgene cassette (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035200#pone-0035200-g001" target="_blank">Figure 1</a>). Three specific bands correlating to genomic DNA, each of the predicted size (4.5, 1.7 and 1.4 kb), are seen in WT samples and all transgenic animals (arrows) and serve as loading control between lanes; the arrowhead indicates the position of the transgene-specific signal at 3.115 kb correlating to a PstI-PstI transgene fragment, which is all-inclusive of the probe. (B) PCR genotyping using a forward primer contained within intron 1 and a reverse primer completely contained within the amelotin cDNA region verifies the generation of a 429 bp amplicon in transgenic mice (lanes 1, 2, 4, 5, 6), but not in non-transgenic littermates (lanes 3, 7). Samples for lanes 1 to 7 were randomly selected from litters from both transgenic lines (57 and 457).