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. Author manuscript; available in PMC: 2017 Jan 28.
Published in final edited form as: Genesis. 2015 Aug 10;53(9):612–626. doi: 10.1002/dvg.22879

Figure 2. Generation of Evc2/Limbin conditional mutant mouse.

Figure 2

(a) A loxP site followed by a Pgk-neo cassette flanked by FRT sites was inserted into intron 12. Another loxP site with a KpnI and a HindIII sites was inserted into intron 14. Positions of 5’ and 3’ external probes for Southern analyses and the sizes of the restriction fragments detected by these probes are shown. Hd, HindIII; K, KpnI. (b) Genomic DNA from wild type ES cells (lane WT), randomly inserted clone (lane 0) and correctly targeted clones (lane 1–3) were digested with enzymes and blots were hybridized with the probes as indicated. (c) Primers E and F were used to differentiate the targeted allele (fn, floxed allele with the neo cassette, or fx, floxed allele without the neo cassette) from WT (top). Presence of the Pgk-neo cassette in the fn allele was confirmed by primers A and B (middle). Primers A and D were used to detect Cre-dependent recombination (dE, Cre recombined allele, bottom). Mixture of the primers A, E and D can differentiate WT, floxed (fn or fx), and the Cre recombined alleles (bottom). The position of primer D is similar to that of primer F, but primer F did not work with primer A. M, 100 bp ladder; *, 600 bp.