Supplemental Material for Kottler et al., 2020 Verena A. Kottler Romain Feron Indrajit Nanda Christophe Klopp Kang Du Susanne Kneitz Frederik Helmprobst Dunja K. Lamatsch Céline Lopez-Roques Jerôme Lluch Laurent Journot Hugues Parrinello Yann Guiguen Manfred Schartl 10.25386/genetics.7957376.v1 https://gsajournals.figshare.com/articles/dataset/Supplemental_Material_for_Kottler_et_al_2020/7957376 Figure S1 contains pictures of melanic and non-melanic <i>G. holbrooki</i>. Figure S2 presents the results of the RAD-sequencing analysis. Figure S3 represents a molecular phylogeny of fish <i>sox9</i>. Figure S4 are alignments of male <i>G. holbrooki </i><i>gipc1 </i>and <i>sox9b </i>alleles and protein sequences. Figure S5 are multiple species alignments of Gipc1 and Sox9 protein sequences. Figure S6 contains PCR results demonstrating that <i>sox9bY </i>was generated by allelic diversification. Table S1 gives information on the fish used for pool sequencing. Table S2 contains the annotated loci within the 100 kb windows with the highest density of sex-specific SNPs. Table S3 presents the results of the allele-specific PCRs. Table S4 contains the sex chromosome genotypes of F<sub>1 </sub>fish (pedigree WLC20074) from a cross between a <i>G. affinis </i>female (WLC1313) and a <i>G. holbrooki </i>male (WLC2514) determined by PCR. Table S5 gives information on the number of offspring of non-melanic females and non-melanic males derived from melanic strains. 2019-11-25 20:13:43 mosquitofish sex determination pool sequencing gipc1 sox9b Developmental Genetics (incl. Sex Determination)