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. 2011 Jul 7;39(18):e121. doi: 10.1093/nar/gkr550

Figure 3.

Figure 3.

Generation of Rnf12 knockout ES cell lines by insertion of a transcription stop cassette. (A) Generation of Rnf12 knockout ES cell lines by integration of a splice acceptor triple poly-adenylation sequence (SA-tpA) together with a kanamycin/neomycin resistance cassette. Different targeting constructs were generated to target the stop cassette to one of the introns 2, 3 or 4. (B) RT–PCR analysis with cDNA form clones targeted with cassettes targeting different introns, using primers amplifying the NheI RFLP located in exon 5. RT–PCR products were digested with NheI to reveal allele specific PCR products. Correctly targeted clones are indicated (ko). (C) Table summarizing the targeting efficiency of the constructs described for targeting the C57BL/6 allele (n = number of clones picked). (Asterisk) Targeting efficiencies of previously described experiments involving C57BL/6, and Cast/Ei exon 5 targeting constructs used to target the 129/Sv and Cast/Ei alleles in F1 2-1 129/Sv/Cast/Ei female ES cells (10,16). (Double asterisk) The Cast/Ei allele was targeted in Rnf12+/− (129/Sv/Cast/Ei) ES cells creating a Rnf12 null ES cell, which may affect the targeting efficiency.