In situ capping is a management technique for contaminated sediments involving the placement of clean material at the sediment-water interface. This work combined porewater geochemical profiling with quantitative microbial data to investigate the intrinsic microbial colonization of a sand cap. Geochemical characterization using voltammetric microelectrodes indicated vertical stratification of biogeochemical processes within a capped sediment column. Following dissection of the column, quantitative real-time PCR (qPCR) enumerated microbial populations within each discrete redoxzone and was accompanied by terminal-restriction fragment length polymorphism (T-RFLP) to elucidate general community shifts. Bacteria and Archaea were present within the cap according to qPCR, with higher concentrations generally observed in the underlying sediment. Iron-reducing populations were detected and quantified using newly designed qPCR primer pairs for Anaeromyxobacter spp. and Shewanella spp. and published primer sets for delta-Proteobacteria and Geobacteracea. Results confirmed geochemical measurements indicating that microbial Fe(III) reduction was a major process in the overlying cap. Genes encoding microbial sulfate reduction (dsrA) and methanogenesis (mcrA) were also present within the cap but were more prevalent in the sediment. Canonical correspondence analysis of terminal-restriction fragment length polymorphism (T-RFLP) patterns verified that spatial changes in bacterial community composition were significantly correlated to depth and Fe2+ and Mn2+ concentration gradients. Cumulatively, results demonstrate that microorganisms indigenous to aquatic sediments colonized the overlying cap to form complex communities mirroring redox stratification. Implications of capping for biogeochemical cycling, contaminant fate and transport, and remedial design are discussed.