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. 2017 Mar 7;7:43883. doi: 10.1038/srep43883

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Agarose gel electrophoresis of control PCR products to confirm correct 3xP3-AmCyan excision via (a) loxN recombination or (b) PUb-AmCyan excision via lox2272 recombination. AH445-F4, M2, -M8, -M9 and -M10 denote tested pools/families. AH0513-M8 is the parental line serving as a control for no excision events. (a) In case of correct loxN excision an 840 bp product is expected in the positive pools, but no product in the parental control due to a large product size (>5600 bp). (b) In case of correct lox2272 excision a 2100 bp product is expected in the positive pools, but no product in the parental control (>8600 bp). To prove that a missing positive PCR band indeed was due to lack of excision, a no excision control PCR was done that would only produce a product if excision did not occur (shown here only for lox2272). See Supplementary Fig. S1 for the complete dataset. NTC represents the no template control reaction for the different primer combinations. Marker denotes the 2-log ladder from NEB. bp = base pairs.