title: 16S: Alterations to grassland soil nutrient supply and plant consumer manipulations jointly influence fungal necromass decay
design: We analyzed microbial communities in soil and on necromass collected at 6- and 32-days post-burial, representing early and intermediate decay stages. For these molecular-based analyses, total genomic DNA was extracted using a Qiagen DNeasy PowerSoil Kit (Qiagen, Benelux BV), including a 30-second bead beating step with 2-mm sterile glass beads (BioSpec Products, Bartlesville, OK, USA) to homogenize the necromass before cell lysis. To assess the composition of the microbial decomposer communities, 16S (bacterial) DNA amplicons were generated at the University of Minnesota Genomics Center (UMGC, Minneapolis, MN). 16S amplicons were generated using a dual-indexed PCR method optimized by the UMGC to amplify the V4 region (primer pair: 515F-806R) with KAPA HiFi polymerase. For PCR, 25 cycles were used for amplicon generation. Samples were sequenced on a full NextSeq run (2  300 bp Illumina chemistry).