Despite historical increases in the number of individuals engaging in same-sex relations and entering same-sex unions, the causes of sexual orientation remain an open question. Two biological processes that have received some degree of empirical validation are the fraternal birth-order effect (FBOE) and the female-fecundity effect (FFE). Respectively, these processes posit that having a greater number of older brothers and being part of larger sibships independently increase the odds of male homosexuality. Nevertheless, previous studies have relied on suboptimal data and methods, including underpowered and selected samples, and models that fail to fully disentangle the two processes. In addition, they have rarely analyzed samples of women. We address these limitations using high-quality, population-level linked register data from the Netherlands (n = 9,073,496). Applying a novel multivariable approach, we jointly examine the FBOE and FFE by comparing the sibship characteristics of men (n = 26,542) and women (n = 33,534) who entered a same-sex union against those who did not (n = 4,607,785 men and 4,405,635 women). Our analyses yield robust evidence of an FBOE on both male and female homosexuality, but no support for the FFE. Additionally, we find that individuals' birth order affects the probability of entering a same-sex union, regardless of the sex of older siblings.