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. 2017 Mar 3;7:59. doi: 10.1038/s41598-017-00140-9

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© The Author(s) 2017

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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Creation of mutant mice with a 2-Mb deletion with a donor plasmid. (A) Schematic representation of the 2-Mb region upstream of Sox9. The black line represents genome sequence. Homology arms are indicated with black and gray boxes. PCR primers are indicated with arrows. Positions of sgRNA recognition sequences are indicated with scissors. (B) An agarose gel electrophoretogram image of PCR products amplified using 2F1 and 1R1 primers and DNA prepared from #48 embryo. M: a 1-kb DNA ladder (molecular weight markers); N: no template control. (C) Sequence of the deletion junction. Sequences of the donor plasmid and of #48 embryo are aligned. Positions corresponding to arms and the ClaI recognition sequence are indicated at the top. Parts of gRNA recognition sequences are underlined. Black and gray boxes correspond to those of (A).

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