Aquifers are rich in microbial diversity. However, there is a lack of information about sessile communities in these environments because of the difficulty in sampling fresh in situ rock surfaces. Thus, this study’s objective was to better understand the sessile community in a fractured aquifer. Additionally, the impact of the rock mineral composition on microbial community composition during colonization was explored. Using a system of bioreactors, we recreated the environmental conditions of a 1.5 m deep aquifer in Covey Hill (QC, Canada) using groundwater samples collected from the site. We carried out 16S/18S rRNA amplicon sequencing of the water and sessile communities after 24 days of incubation. Our data showed that many microbial taxa overlapped between the sessile and planktonic communities, indicating colonization of the solid surfaces. Quartz and feldspar had a significant impact on bacterial community structure. Sessile communities were dominated by Gaillonella, Alkanindiges, unclassified Acetobacteraceae, Apoikiales, Glissomonadida, and Synurales. We could not detect any Archaea in the sessile community. The sessile communities contained bacterial genera involved in iron cycling and adapted to acidic and low-carbon-concentration environments. Eukaryotic predators dominated the sessile community.