Schematics for Col1a1-directed expression of FRT-Cre-ERT2-FRT and FRT-STOP-FRT-Cre-ERT2, the removal of the neomycin selection cassette, and Southern blot.

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Schematics for Col1a1-directed expression of FRT-Cre-ERT2-FRT and FRT-STOP-FRT-Cre-ERT2, the removal of the neomycin selection cassette, and Southern blot for successful homologous recombination. Schematics for Col1a1-directed expression of FRT-Cre-ERT2-FRT and FRT-STOP-FRT-Cre-ERT2, the removal of the neomycin selection cassette, and Southern blot for successful homologous recombination. Schematic of the Col1a1FRT-Cre-ER-T2-FRT (A) and Col1a1FRT-STOP-FRT-Cre-ER-T2 (B) alleles. The neomycin cassette is used for selection of ES cells with integration of the targeting construct. PhiC31 integrase is used to remove the neomycin selection cassette in vivo after germline transmission. Locations of genotyping primers are indicated on each schematic. (C) Southern blot analysis indicates successful homologous recombination of the Col1a1FRT-Cre-ER-T2-FRT-NEO allele in the E2, E1 and B7 clones. The B7 clone was used to generate the Col1a1FRT-Cre-ER-T2-FRT-NEO line. (D) Southern blot analysis indicates successful homologous recombination of the Col1a1FRT-STOP-FRT-Cre-ER-T2-NEO allele in the E1, C1 and B11 clones. The B11 clone was used to generate the Col1a1FRT-STOP-FRT-Cre-ER-T2-NEO line. Minsi Zhang, and David G. Kirsch Dis. Model. Mech. 2015;8:1155-1166 © 2015. Published by The Company of Biologists Ltd