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. 2010 Jan 13;38(7):e96. doi: 10.1093/nar/gkp1234

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Gene-replacement strategy. (A) Scheme for gene replacement. Cre recombinase-mediated integration regenerates a functional hygromycin resistance gene while also resulting in loss of neomycin resistance. The dashed line indicates the expected fragment size after HincII digestion in the Southern blotting. Arrows (P5–P8) are PCR primers used below. (B) Southern blotting results for one hygromycin-resistant clone using an EGFP probe. M: Size marker, R: Clone that underwent gene replacement, P: Parental clone. (C) PCR genotyping of the hygromycin-resistant clone. (D) EGFP expression of the hygromycin-resistant clones. (E) Neomycin sensitivity of hygromycin-resistant and EGFP-expressing clones. (F) EB formation of the hygromycin-resistant clones. (G) Neural differentiation of a hygromycin-resistant clone.