Host DNA contamination impedes the molecular analyses of microbiomes in host samples, such as bodily fluids and swabs. The SPINeasy Host Depletion Microbial DNA Kit provides an easy-to-use workflow to isolate microbial DNA from samples containing high amounts of host DNA. This background reduction of host DNA is achieved through selective lysis of host cells with our specially formulated Host Lysis Buffer. Host DNA, which has been released in the solution, is then removed from the supernatant through centrifugation. A further host depletion treatment provides enzymatic degradation of any remaining host DNA, as well as DNA from dead microbial cells. Finally, the remaining intact cells are lysed through a combination of specially formulated buffers and highly efficient homogenization using FastPrep instruments from MP Biomedicals. Microbial DNA is purified using a convenient silica-membrane spin-column technology workflow and ready for downstream molecular applications.