The ratio of the lengths of the second and fourth fingers (2D:4D) may serve as a marker for prenatal androgen signaling. Because people are typically unaware of their 2D:4D, its use allows possible effects of early sex hormone regimes and socialization to be disentangled. We conducted a meta-analysis on relationships between 2D:4D and sexual orientation in men and women in 18 independent samples of men and 16 independent samples of women. Collectively, these samples comprised 1,618 heterosexual men, 1,693 heterosexual women, 1,503 gay men, and 1,014 lesbians. In addition to identifying the normative heterosexual sex difference in 2D:4D for both hands, we found that heterosexual women had higher (more feminine) left- and right-hand 2D:4D than did lesbians, but we found no difference between heterosexual and gay men. Moderator analyses suggested that ethnicity explained some between-studies variation in men. These results add to a literature suggesting that early sex hormone signaling affects sexual orientation in women, and highlight the need for further research exploring the relationships among 2D:4D, sexual orientation, and ethnicity in men.