The present study developed protocols to exploring the different reproductive behaviors between urban and rural Gryllus pennsylvanicus populations. This study uses a unique approach to estimating mating rates among urban and rural crickets by genotyping the sperm stored in a field-caught female’s reproductive tract and using this to estimate the number of males she mated with. I examined the efficacy of four microsatellite loci for distinguishing individuals in the field cricket species Gryllus pennsylvanicus and established a protocol for extracting and genotyping sperm DNA stored in the female reproductive tract. I found that loci PG1,G3, Gr143, and G28 were effective makers for genotyping Denver G. pennsylvanicus and could be used to distinguish each individual in my study. Furthermore, I established a sperm DNA extraction protocol that will allow future undergraduate students to explore the mating behavior of Gryllus pennsylvanicus.