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Fig. S3 RT-PCR confirms indel mutations and identifies aberrant splice variants caused by mutations.
(A) RT-PCR products for all gene mutants along with WT control. The amplified products were resolved on 2% agarose gel. RT-PCR was designed to amplify the exon containing the indel mutation in knockouts and wild-type fish. Minus RT control was performed for hg50 and hg51 lines due to fancf being a single exon gene. Expected size products were observed for all mutants, except hg41 (fanca), hg42 (fancb), hg45 (fancd1), and hg58 (fancl) mutants, as denoted by the red arrows. Amplicons were sequenced to confirm the mutation, and to determine any aberrant splicing. Multiple products were sequenced after cloning into a vector. (B-E) Representative chromatograms for aberrant splice products of fancl_hg58 (B), fanca_hg41 (C), fancb_hg42 (D), and fancd1_hg45 (E). RT-PCR primers for actb2 were used as transcript control.