Female crayfish stores male gametes after mating until the beginning of egg laying and fertilization. The aim of the present study was to investigate the duration of post-mating spermatophore storage as well as the timing and temperature of spawning in two crayfish species of economic importance, namely the signal crayfish Pacifastacus leniusculus and the noble crayfish Astacus astacus. Results showed that the average duration of the post-mating spermatophore storage is significantly (P<0.05) longer in the noble crayfish (34.6±1.7 days, range: 19 to 60 days) than the signal crayfish (3.9±0.5 days, range: 1 to 18 days). The highest percentages of the post-mating spermatophore storage duration in the signal crayfish (46.5%) and the noble crayfish (44.5%) were 1 and 31 to 40 days, respectively. While there is an overlap in the timings of mating and egg laying in the signal crayfish, these two reproductive processes were not observed at the same days in the noble crayfish and there was at least 2 weeks interval between last mating and first egg laying individuals. Average mating and egg laying temperatures were significantly (P<0.05) higher in the signal crayfish than the noble crayfish. The average temperatures for mating in both species were significantly (P<0.05) higher than the temperatures that they utilized for egg laying. In conclusion, female noble crayfish stores post-mating spermatophores a longer duration compared with the signal crayfish. Also, the signal crayfish mates and lays egg in temperatures that are higher than the noble crayfish. Spawning season is shorter in the signal crayfish compared with the noble crayfish. The results of present study provide information contributing to the crayfish broodstock management in aquaculture.