Dataset: reconstructed genomes
Deployment: MSM20-5

Crustal fluids were collected from the single horizon at U1382A and from the shallow, middle and deep horizons in U1383C (Edwards et al., 2012) using a mobile pumping system designed for microbial sampling from CORK fluid delivery lines as described in Meyer et al. (2016) and Cowen et al. (2012). Deployed with the ROV system, mobile pumping system connectors are attached to the CORK wellhead via an umbilical to the hydrological zone of interest within the aquifer.  In 2012, 12 l of each fluid sample were filtered on to a 0.22 μm Sterivex-GP filter (Merck Millipore, Billerica, MA, USA) as described in Meyer et al. (2016). In 2014, 12 l of each sample was filtered in situ and immediately fixed with RNALater (Thermo Fisher Scientific, Waltham, MA, USA), as described previously (Akerman et al., 2013). After sampling in 2012, a battery-powered GeoMICROBE sled was left at each CORK for time series autonomous sampling of the fluid delivery lines (Cowen et al., 2012). For each filter sample, ~10 l of fluid were filtered in situ and immediately fixed with RNALater.  Sleds were deployed in April 2012 and recovered in April 2014 with samples collected. Upon sled recovery, filters were transferred to fresh RNALater and stored at −80 °C, while all bag samples were stored at 4 °C (Cowen et al., 2012). Deep bottom water was sampled in 2012 and 2014 via a CTD at 100 m above the seafloor and filtered in the same manner as the crustal fluids onto Sterivex filters. Total microbial biomass in fluids was enumerated with DAPI (4′,6′-diamidino-2-phenylindole; Sigma-Aldrich, St Louis, MO, USA) and epifluorescent microscopy (Porter and Feig, 1980).