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dc:title: Figure S1
modified: 2013-06-25T17:51:20
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dc:description: Targeting strategy to generate geminin conditional knockout allele. A) Two LoxP sites were inserted in the first and third introns of geminin genomic locus upon site-specific recombination. The floxed allele possesses exon 2 and 3 flanked by LoxP sites and upon Cre mediated recombination exons 2 and 3 are excised. Thus the remaining conditional knockout allele loses its ability to produce functional protein (adapted after [55]). B) Southern blot analysis of the Gmnnfl/+ ESCs showing correct integration by indicated restriction enzymes. C) iGmnn ESCs were cultured for 72 hours in ES-CM and were treated for different periods of time with tamoxifen. The genomic DNA was extracted and genotyped. Different combinations of primers in separate reactions were used to amplify the floxed and recombined knockout alleles. The same amount of genomic DNA was used for each reaction. D) iGmnn ESCs were cultured for 96 hours in ES-CM and were treated for different periods of time with tamoxifen. The amount of geminin in whole cell lysates were anlaysed by western blot band quantified by imageJ.(TIF)
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description: Targeting strategy to generate geminin conditional knockout allele. A) Two LoxP sites were inserted in the first and third introns of geminin genomic locus upon site-specific recombination. The floxed allele possesses exon 2 and 3 flanked by LoxP sites and upon Cre mediated recombination exons 2 and 3 are excised. Thus the remaining conditional knockout allele loses its ability to produce functional protein (adapted after [55]). B) Southern blot analysis of the Gmnnfl/+ ESCs showing correct integration by indicated restriction enzymes. C) iGmnn ESCs were cultured for 72 hours in ES-CM and were treated for different periods of time with tamoxifen. The genomic DNA was extracted and genotyped. Different combinations of primers in separate reactions were used to amplify the floxed and recombined knockout alleles. The same amount of genomic DNA was used for each reaction. D) iGmnn ESCs were cultured for 96 hours in ES-CM and were treated for different periods of time with tamoxifen. The amount of geminin in whole cell lysates were anlaysed by western blot band quantified by imageJ.(TIF)
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dcterms:created: 2013-06-25T17:51:20
dcterms:modified: 2013-06-25T17:51:20
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title: Figure S1
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