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Supplemental figure 1: Sequencing of GFP+ and GFP- colonies confirms that a positive GFP signal is indicative of a wild type or in-frame mutation while the absence of GFP signal indicates an out-of-frame mutation at the TALEN-targeted locus of the genome in animals injected with TALEN mRNAs targeting Xenoups tropicalis Sox3 gene. The mRNAs encoding the left and right arms of a TALEN targeting the Sox3 region shown in Figure 4A were injected into fertilized eggs and the animals were reared into tadpole stages. The genomic DNA was isolated from the tadpoles, PCR amplified, and cloned into pmCherry-GFP construct and analyzed as shown in Figs. 2 and 4. Colonies with red fluorescence of mCherry expression were selected and sorted into GFP-negative (A) and GFP-positive (B) groups, grew in LB medium supplemented with ampicillin. Plasmid DNA was isolated from them and subjected to DNA sequence analysis. TALEN targeted sequences are in blue and the areas are shaded. Note that all the GFP-negative clones contained out-of-frame mutations while the GFP-positive clones were either wild type or in-frame mutants. The mutations with deletions (-) and additions (+) were indicated. When there were more than one colony of a particular mutation, the number of colonies was shown in parenthesis. Image published in: Fu L et al. (2016) Copyright © 2016, The Author(s). This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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