Marker-assisted selection (MAS) use in breeding programs allows for examination of seedlings at an early stage before accumulation of high field costs. However, introducing MAS into a breeding program implies additional costs and uncertainties about effective incorporation. Previous simulations in apple (Malus ×domestica) have shown cost-effective applications of MAS. To further evaluate MAS cost-effectiveness in perennial crops, we conducted a cost-effectiveness analysis examining MAS in an upper midwestern U.S. peach (Prunus persica) breeding program. Breeding program procedures and associated costs were collected and used as input into spreadsheet-based simulations of the breeding program. Simulations compared a conventional breeding program to MAS with varying cull rates of low, medium, and high at multiple stages in the breeding cycle. Cost-effective MAS implementation was identified at the end of seedling trials with a break-even cull rate of 4%. These results inform breeders of cost-effectiveness of MAS use in a peach breeding program.