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. 2010 Aug 11;38(22):8188–8195. doi: 10.1093/nar/gkq707

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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 3. — Construction of the RFzero strains. (A) Genetic modifications to create the RFzero strains. TF and KO mean transformation and knock-out, respectively. (B) PCR amplification was performed, using primers (black arrows) to amplify the entire prfA locus (Lanes 1 and 2) and primers (gray arrows) to amplify the prfA sequence inside the knocked out region (Lanes 3 and 4), with the genomic DNAs of HST08 (Lanes 1 and 3) and RFzero-q (Lanes 2 and 4) as the template. (C) PCR amplification of the prfA sequence inside the knocked out region, with the genomic DNAs of the HST08 strain (Lanes 1, 3, 5, 7, 9, 11 and 13) and the RFzero-q strain (Lanes 2, 4, 6, 8, 10, 12 and 14), each with BAC7 (Lanes 1 and 2), and BAC6 plasmids (Lanes 3–14) lacking the genes indicated above the lanes.