Zooplankton resting egg banks accumulate resting stages of various zooplankton species that are active in different habitats and different periods of the year. As such, hatching of resting eggs from lake sediments may potentially be very useful in zooplankton diversity studies. In this study, we tested whether the efficiency of the cost-effective technique is increased by isolating the resting eggs from the sediment prior to incubation. Isolation of the eggs was advantageous for the overall hatching success (+26% after 36 days of incubation compared to incubation of sediment). Furthermore, isolation of resting eggs makes egg bank diversity analyses less time consuming in two ways. ( 1) It reduced the time needed for the eggs to hatch with on average 35%. In the isolation treatment all responsive resting eggs hatched within the first 4 weeks of incubation, while in the non-isolation treatment neither the cumulative number of macrozooplankton hatchlings nor the cumulative number of hatched cladoceran species levelled off after 36 days of incubation. ( 2) In contrast to the non-isolation treatment, where large differences occurred between taxa in incubation time, isolation reduced such inter-specific differences, so that even very short incubation periods kept bias within acceptable limits.