Supplementary Figure 4: Identification of correctly targeted Ctnnb1 mESC clones.
From: One-step generation of conditional and reversible gene knockouts
(a) The FLIP cassette containing a resistance gene is inserted into the exon 5 of Ctnnb1. SD, splice donor; SA, splice acceptor, BP, branching point; purple triangles, LoxP1 sites; Pink triangles, Lox5171 sites.LoxP It is noteworthy that the LoxP1 and Lox5171 sites cannot recombine with each other but only with LoxP sites of the same sequence.
(b) PCR detection of FLIP cassette insertion in the Ctnnb1 locus. Correctly targeted clones E2 and B1 are positive for 5’ and 3’arm genotyping PCR reactions (for genotyping strategy see Supplementary Fig. 2b). Exon 5 PCR detects the remaining allele. The clones (E2 and B1) are correctly targeted.
