To test the response of microbial communities to periodic oxygen limitation, we conducted a laboratory experiment where two contrasting soils (a rainforest Oxisol from Puerto Rico, and an Iowa cropland Mollisol) were incubated under headspace treatments where oxygen availability varied cyclically over time. Treatments consisted of 0, 2, 4, 8, or 12 d of anoxic conditions (dinitrogen headspace) followed by 4 d of oxic conditions (i.e., ambient oxygen concentrations), and these treatments were repeated for a total of 384 d. At 0, 48, and 384 days, DNA was extracted from replicates from each treatment for sequencing of 16S rRNA amplicons. Companion biogeochemical measurements from this experiment were published previously by Huang et al. (2021a,b). These data support the Hall et al. (2022) manuscript published in Frontiers in Microbiology.