Summary The analyses of haploid male DNA pooled (MDP) samples can increase the cost efficiency in population genetic studies for a variety of applications in fundamental and applied screening of social Hymenoptera populations. Depending on the research problem addressed it can reduce the number of genotyped samples per study by orders of magnitude. In this study we show the feasibility of MDPs for assessing population allele frequencies, mother queen genotypes and mutation rates for microsatellite loci in the honeybee (Apis mellifera). The main purpose of MDP genotyping is not to replace individual based analyses but to discard non-informative samples and monomorphic loci from further analyses.