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Fig. 3 Detection of tjp1a-mRFP integration and germline transmission. (a) Confocal image of the lens of a 3 dpf embryo injected with the tjp1a-mRFP targeting template expressing mRFP. mRFP is observed at the border of cells within the lens and some surrounding structures. E designates the eye. (b) 1.2% agarose gel displaying junction fragments from a PCR run on fin clips of F1 tjp1a-mRFP is86 embryos expressing mRFP. Bands are seen at approximately the expected sizes of 270 bp and 230 bp. (c) Annotated Sanger sequencing results from sequencing of the bands shown in (b). The junction sequences recovered were color coded and compared to the reference sequence: grey, gene specific primer; black, mRFP primer; red, tjp1a gene sequence; blue, homology arms; green, mRFP sequence.