title: 16S amplicon of Soil
design: Sequencing libraries of bacterial and fungal communities of soil (MP16 treated and non-treated) were prepared according to the Illumina 16S Metagenomic Sequencing Library Preparation guide. For the bacterial community, the V3-V4 regions of the 16S bacterial rRNA gene were amplified using a two-step PCR protocol with V3-V4 primers (For: 5-CCTACGGGNGGCWGCAG-3, Rev: 5- GACTACHVGGGTATCTAATCC-3).Sample-specific 6-bp barcodes were incorporated into the primers for multiplex sequencing. The PCR reaction was done using the Phusion High-Fidelity PCR Master Mix (NEB). The PCR products were verified on 2% agarose gel, and 400-450 bp bands for 16S were cut and mixed in an equal-density ratio and then purified the PCR product with Qiagen Gel Extraction Kit (Qiagen). Following the manufacturer's recommendations, the generated sequencing libraries using NEB Next Ultra DNA Library Prep Kit for Illumina added the index codes. The library quality assisted using the Qubit dsDNA high-sensitivity (HS) kit (Thermo-Fischer Scientific, Schwerte, Germany) and Agilent Bioanalyzer 2100 system. The library was normalized and sequenced on the Illumina platform (Illumina Inc., San Diego, CA, USA) and generated 2x 250 bp paired-end reads.