Zooplankton in temporary waters produces resting stages to survive recurrent dry periods. Branchiopod crustaceans (i.e., cladocerans, large branchiopods) overcome these periods in the form of resting eggs buried in the sediment. Examining the diversity in the resting egg banks allows for a more accurate estimation of biodiversity of aquatic ecosystems than looking only at the active communities. The isolation of resting eggs from the sediment may be achieved by the sugar flotation method, which usually results in higher density and diversity than untreated samples (i.e., incubated in the sediment). We tested the effect of sugar isolation and centrifugation on the hatching success of resting eggs already isolated from sediment in order to reveal any direct effects on hatching success. We used four different branchiopod species, Daphnia magna, Moina brachiata, Branchinecta orientalis, and Triops cancriformis. Although we hypothesised that osmotic stress caused by sugar and centrifuging influence the hatching success either positively (e.g., faster activation as a response to osmotic changes) or negatively (destroyed by centrifugation), we found no significant difference either in the timing or rate of hatching between centrifuged and non-centrifuged eggs. Once the eggs are exposed to light and/or oxygen availability by being removed from the sediment, the centrifugation process does not have any additional effect on their hatching. Regardless of treatment, we found a significant difference in the hatching timing in the two major groups, with large branchiopods hatching earlier than cladocerans. We found that the sugar flotation method itself does not influence the hatching fraction of branchiopod resting eggs (implying no adverse effect on their viability) and its success in enhancing hatching can be attributed to its efficiency in removing eggs from the sediment.