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. 2016 Dec 20;7:1904. doi: 10.3389/fpls.2016.01904

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Copyright © 2016 Malnoy, Viola, Jung, Koo, Kim, Kim, Velasco and Nagamangala Kanchiswamy.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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CRISPR/Cas9 RNPs in vitro digestion assay results of each sgRNAs used in the study. (A) In vitro digestion of targeted loci at MLO-7 gene. (B) In vitro digestion of targeted locus in the DIPM-2 gene. (C) In vitro digestion of targeted locus in the DIPM-2 gene. (D) In vitro digestion of targeted locus in the DIPM-4 gene. In each group, non-treated (sgRNA: x; Cas9: x) or Cas9-only (sgRNA: x; Cas9: o) samples were used as a control. After treatment with Cas9 with targeted sgRNA (RG1 to RG8 in each group) amplified target genomic DNA was digested and smaller bands were detected in gels after electrophoresis. Groups showing an intense band after digestion (indicated with black boxes) were selected and used for further experiments.