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. 2015 Mar 16;10(3):e0119818. doi: 10.1371/journal.pone.0119818

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© 2015 Tominaga 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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Fig 4 — (a) Killing efficiency of dP-selection against E. coli strain harboring a single hsvtk gene (HK cassette) or duplicated hsvtk (HKH cassette), ND: Not detected. The bar heights show the average of 3 samples, and error bars indicate the standard deviation. (b) The experimental workflow for a two-round consecutive recombineering: lacZ and proV loci were serially replaced with pL-gfp ^mut3.1 and pT5-mrfp cassette respectively using HK or HKH. All DNA cassettes used here were prepared with 40-bp homology arms to the individual target sites, except the step for replacing lacZ with pL-gfp ^mut3.1 cassette where 1-kbp homology arms were adopted.