16S Porewater: SPRUCE
Porewater samples were keep on ice before being filtered in a field lab and stored at -80C. DNA was extracted from porewater filters using the Qiagen PowerSoil DNA Kit according to the manufacturers instructions. DNA quality was assessed with a Nanodrop spectrophotometer, quantified using a Qubit High-Sensitivity (HS) assay, and verified for integrity on an agarose gel. Samples were sequenced at Georgia Institute of Technology using primers 515F-Y (5-GTGYCAGCMGCCGCGGTAA-3) and 806R-Apprill (5-GGACTACNVGGGTWTCTAAT-3) with overhang adapters (Apprill et al. 2015; Caporaso et al. 2011). Barcoded amplicons were sequenced on an Illumina MiSeq2000 platform using a 500-cycle v2 kit to generate paired-end reads of 250 bp.