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. 2012 Sep 14;7(9):e44974. doi: 10.1371/journal.pone.0044974

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© 2012 Buck et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Restriction digestion of 1^st stage and 2^nd stage recombinant BAC clones to confirm the expected pattern changes and exclude unwanted rearrangements. A. BamHI and EcoRV restriction digestion. Lane 1. Unmodified pBAC_MMA, Lane 2. 1^st stage recombination (insertion of the I-SceI/kanR cassette), Lane 3. pBAC_MMA*, R403stop (2^nd stage of recombination). B. EcoRV restriction digestion of the 1^st stage of recombination (Lane 2) showing loss of the 4.7 kb band with the insertion of the I-SceI/kanR cassette (appearance of the expected 6.8 kb band). The restriction digestion pattern of the unmodified pBAC_MMA (Lane 1) and the pBAC_MMA*, R403stop (2^nd stage of recombination) (Lane 3) are identical.