Plasmids coding TALE nucleases were constructed using the Platinum Gate TALEN Kit (1000000043: Addgene, Cambridge, MA, USA)26 (link). mRNAs coding the TALE nucleases were synthesised using mMESSAGE mMACHINE T7 Transcription kit (AM1344; Thermo Fisher Scientific) and injected into dechorionated unfertilised eggs with a glass capillary at the concentration of 150 ng/μL each with rhodamine dextran as an injection marker (Fig. S1e ).
The mutagenic efficiency of TALEN pairs was examined by amplifying the target regions from the genomic DNA extracted from a mixture of 10 tailbud larvae with primers shown in Supplementary Table4 , cloning the products with TOPO TA Cloning Kit (450641; Thermo Fisher Scientific), performing colony PCR with M13For and M13Rev primers, and analysing the sequence of the gel exudates after agarose gel electrophoresis using BigDye Terminator v3.1 Cycle Sequencing Kit.
The mutagenic efficiency of TALEN pairs was examined by amplifying the target regions from the genomic DNA extracted from a mixture of 10 tailbud larvae with primers shown in Supplementary Table
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