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. 2010 Aug 26;17(5):293–301. doi: 10.1093/dnares/dsq020

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© The Author 2010. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.

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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Insertion of the loxP cassette into the tet-O HAC by homologous recombination in DT40 cells. (A) Morphology of A9BH-3 hybrid cells. (B) Genomic DNA from A9BH-3 hybrid cells was analysed by PCR using HAC-specific primers, HRL-F/R and BACpr-F/R. (C) FISH of DT40/BH-1 cells using the hCot1 probe. The tet-O HAC is indicated by an arrowhead. (D) Insertion of the loxP/HPRT cassette into the tet-O HAC using the targeting vector 5–4–3, which carries the regions of homology (black boxes) to BAC vector sequences in the HAC. (E) FISH analysis of DT40/BHI 1–38 (left) and DT40/BHI 2–2 (right) cells. FISH analysis was performed using a biotin-labelled probe for the loxP cassette (green) and digoxigenin-labelled human hCot1 (red).