Soil Microbial DNA Extraction and Sequencing

Soil DNA was extracted according to Mandakovic et al. (2018a (link)) with some modifications. Total DNA from soils was extracted from 10 g of each sample using the NucleoSpin® Food kit (Macherey-Nagel), following the manufacturer's instructions, and the CTAB extraction buffer published by Zhou et al. (1996 (link)). Microbial 16S rRNA genes were amplified according to Mandakovic et al. (2018b (link)) without modifications. DNA libraries were constructed following the TruSeq DNA sample preparation (Illumina, USA) protocol. Sequencing was performed by MrDNA Next Generation Sequencing Service Provider (Shallowater, Texas, USA) on an Illumina MiSeq platform in an overlapping 2 × 300 bp configuration, to obtain a minimum throughput of 40,000 sequences (reads) per sample.