A 10.6 m female whale shark Rhincodon typus caught off the coast of eastern Taiwan in 1995 carried 304 embryos that ranged in developmental stage from individuals still in egg cases to hatched and free-swimming near-term animals. This litter established that whale sharks develop by aplacental yolk-sac viviparity, with embryos hatching from eggs within the female. The range of developmental stages in this litter suggested ongoing fertilization over an extended period of time, with embryos of different ages possibly sired by different males. A series of 9 microsatellite markers for R. typus have now been used to investigate paternity in a subset of these embryos. We determined the paternity of 29 embryos representing 10% of the original litter, and spanning most of the range of size and developmental stage of the 304 embryos. All were full siblings sired by the same male, suggesting that this male may have sired the entire litter. Probability analysis indicates that a second male could go undetected if it sired less than 10% of the litter. The range of developmental stages of embryos from this single sire further suggests that female whale sharks may have the ability to store sperm for later fertilization. In the absence of any tissue to determine parental genotypes, maternal mitochondrial sequence was obtained from the embryos, identifying a novel haplotype linked to those from the western Indian Ocean. This finding adds further support for the global population structure emerging for R. typus.