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. 2014 Oct 3;9(10):e109196. doi: 10.1371/journal.pone.0109196

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© 2014 Bruun 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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A Schematic overview of the new pQL11 plasmid B HEK293T cells were infected at low MOI (0,1) of “linked” pQL11 based chimeric Env/V3-model-library viruses and were applied to a FACS-panning procedure 48 h after infection (as described in Material and Methods and Figure 1/5). Env genes were amplified from genomic DNA of collected cells, cloned into fresh pQL11 and were analyzed by qPCR. New virus batches were produced with the resulting new pQL11 Env-library mixtures for infections of fresh cells, thus entering another cycle of selection. The mean values of three independent experiments are shown. Statistics were calculated using the 1-way-ANOVA- (testing, whether mean values differ) and “Dunnett's” post-test (testing, which mean values differ: * P<0.05; ** P<0.01; *** P<0.001). The relative amounts per variant of the input mixture, 1^st and 2^nd cycle are shown as analyzed by qPCR (Efficiency∧^-Ct as % of each variant). The total amounts of pQL11 Env/V3-plasmids in every sample was tested by a reference primer pair that amplifies a sequence independent of the Env genes, respectively (data not shown).