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. 2012 Aug 1;7(8):e42641. doi: 10.1371/journal.pone.0042641

Figure 1. Representation of the construction and validation of the Pitx3-Cre knock-out/knock-in.

Figure 1

A) Schematic drawing of the targeting construct design. B) Schematic representation of the construct insertion validation. Primers (red arrows) are chosen as follows: in the 5- flanking region and in the En2 splice acceptor (5-arm-junction-PCR); in the 3′ flanking region and in the Hygo TK sequence (3′-arm-junction-PCR); in the miCRE sequence (Internal-PCR) C) Validation of homologous recombination through PCR analysis using primers outside the construct at the 5′- and 3′ arm and an internal control (miCre). 1–4: Pitx3 exons.; Open squares indicate 5′ and 3′ Utrs; Closed squares indicate coding region.; mEn2 Spl. Acc: mouse En2 Splice acceptor sequence; *: positive Pcr product in genotyping.