Robertsonian translocation (RT) is a common cause for male infertility, recurrent pregnancy loss, and birth defects. Studying meiotic recombination in RT‐carrier patients helps decipher the mechanism and improve the clinical management of infertility and birth defects caused by RT. Here we present a new method to study spermatogenesis on a single‐gamete basis from two RT carriers. By using a combined single‐cell whole‐genome amplification and sequencing protocol, we comprehensively profiled the chromosomal copy number of 88 single sperms from two RT‐carrier patients. With the profiled information, chromosomal aberrations were identified on a whole‐genome, per‐sperm basis. We found that the previously reported interchromosomal effect might not exist with RT carriers. It is suggested that single‐cell genome sequencing enables comprehensive chromosomal aneuploidy screening and provides a powerful tool for studying gamete generation from patients carrying chromosomal diseases.