Site A1, spring, 49.5m N transect
Soil samples were collected from two sampling sites, A1 (68deg20'35.8"N 18deg50'34.2"E) and A2 (68deg20'47.6"N, 18deg49'24.71"E). Four replicate 50 m transects were established, each at 90 deg from a randomly chosen origin point. Soil cores were collected with a 2.5 cm diameter corer, to approximately 12 cm depth. Cores were stored in the dark in sterile 50 mL polypropylene tubes and kept chilled for a maximum of 2 h before freezing at -20 degC. Frozen cores were transported in insulated containers back to Aberystwyth University. Community genomic DNA was extracted from 100 mg of freeze-dried soil by bead-beating for 30 seconds with four 3 mm diameter sterile glass beads in a Mini8-beadbeater using the phenol-chloroform-cetyl-trimethylammonium bromide protocol of Griffiths et al. (2000) with polyethylene glycol-6000 nucleic acid precipitation for 2 h. Samples collected at a distance of 49.5m along each transect were selected for multiplexed amplicon semiconductor sequencing of the V1-V2 region of the bacterial 16S rRNA gene defined by primers B-27F + MID and A1-357R using the Ion Torrent platform.
Aberystwyth University
AbiSpace_1MN495