The dramatic variation of cichlid fish colour pattern is thought to function in mate choice, evolve by sexual selection, and contribute to explosive speciation. Here, we combine linkage mapping and population genetic analyses to identify a single region of the cichlid genome responsible for the orange blotch (OB) colour phenotype. In each analysis, OB is tightly linked to the c-ski1 gene. Additionally, we use comparative mapping information from the Takifugu rubripes and human genomes to suggest positional candidate loci for OB. Our work should engender a more comprehensive understanding of the molecular ecology of OB and its role in cichlid speciation. Moreover, we have assembled the components of a method to focus upon the genetic basis of evolutionarily and ecologically significant phenotypes.