Xenbase Image ID: 153243

Supplemental figure 2: Detection of out-of-frame mutations induced by four other TALENs with a combination of X-gal staining and GFP fluorescence. B) PCR fragments flanking the TALEN targeting sites in Sox3, TRα-LBD, or Dot1L (Dot1L-TALEN1 and Dot1L-TALEN2) genes were amplified with the NestF(BamHI) and NestR(EcoRI) from genomic DNA isolated from a wild type tadpole (Wt) and F0 TALEN-injected tadpoles (M). The amplified fragments were double-digested with BamHI and EcoRI followed by calf intesitinal alkaline phosphatase treatment, and cloned into the BamHI and EcoRI double-digested pLacZα-GFP as described in Fig.5. The resulting plasmids were transformed into bacteria. The bacterial plates were stained with X-gal. Pictures were taken under the bright field (Bright Field) to visualize X-gal staining (blue) or under a fluorescent microscope for GFP. Note that the colonies from plasmid containing Wt DNA at different loci were uniformly greenish under the bright field, likely reflecting the blue color of the X-gal staining plus weak green color due to the GFP under visible light. On the other hand, colonies from the mutant tadpole (M) were a mixture of blue (lack of GFP) or greenish colonies in the bright field. The greenish colonies for both Wt and mutant animals had strong GFP signal under a fluorescent microscope for GFP. Three independent samples from TALENs-injected tadpoles were analyzed for each TALEN target and a representative picture is shown in the figure. The experiments were repeated twice. 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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