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Schematic outline of the mutant medaka library generation and screening. Male G0 fish were ENU-mutagenized and crossed with wild-type (WT) females. Male F1 progeny were used for sperm cryopreservation and parallel DNA isolation. The library was screened for induced mutations in target genes of interest by dideoxy resequencing. Interesting mutants were retrieved from the cryopreserved archive by in vitro fertilization and incrossed to homozygosity for phenotypic analysis.
