This data release includes all of the data presented in the peer-reviewed publication “Life at the frozen limit: Microbial Carbon Metabolism Across a Late Pleistocene Permafrost Chronosequence”. We collected permafrost from a Pleistocene chronosequence (19 ka to 33 ka) to examine (1) changes in the functional genetic potential of extant microbial communities to metabolize polysaccharides, (2) shifts in the quantity and quality of anions and dissolved nitrogen, and (3) changes in the molecular composition of dissolved organic matter. The data released herein shows that the age of permafrost had a marked effect on both the molecular composition of dissolved OC and the microbial community. Here we clearly demonstrate that while paleoclimate and paelovegetation affects the initial composition of permafrost carbon, microbes metabolize the carbon over time altering its characteristics.