Control Biopile T0 Replicate 1
Soil samples were homogenized in sterile 0.85% NaCl solution and subjected to sequential centrifugation steps to remove soil particles and recover microbial cells. The resulting pellet was resuspended in phosphate-buffered saline (PBS), and total DNA was extracted in triplicate using a commercial soil DNA extraction kit following the manufacturers protocol, with DNA concentration measured by fluorometry. The bacterial 16S rRNA gene (V3V4 region) was amplified using primers 341F (5-CCTACGGGNGGCWGCAG-3) and 785R (5-GACTACHVGGGTATCTAATCC-3), and sequenced on an Illumina MiSeq platform to generate paired-end reads, including technical replicates for each sample.