title: B4A42
organism: mixed sample
description: Protocols: On 10 June 2022, during peak biomass season, we took six soil cores from each of the 48 plots using a 1-cm-diameter soil auger (each 10 cm deep). We did this in all 4 corners (each 30 cm from the edges of the plot) and twice in the center (50 cm from south to north edge and either 40 cm from west or east edge). Soil samples per plot were combined in a plastic bag and stored in a cool box until transported to the laboratory. Samples were sieved using a 2-cm mesh sieve to remove stones, plant material and larger animals. A representative subsample of the sieved soil from each plot was frozen at -20°C for later DNA analyses, the rest was stored at 4°C for soil property analyses. DNA was extracted from 1 g of soil according to Harkes et al. (2025). Sequencing libraries were generated by pooling PCR products of the samples in equimolar ratio's to ensure even sequencing depth. The libraries were then cleaned using NucleoMag NGS Clean-up and Size Select beads (0.6X bead:sample ratio) to discard unwanted contaminants and short DNA fragments (<600 bp). Of each library, 150 fmol was prepared for nanopore sequencing using kit SQK-LSK112 (Oxford Nanopore Technologies Plc., UK) following instructions of the manufacturer.