title: Nanopore shotgun metagenomic sequencing of BS pool
design: To simultaneously isolate RNA and DNA from soil samples, the PowerSoil total RNA kit (Qiagen) and PowerSoil DNA elution kit (Qiagen) was used following the manufacturer's recommendations with minor modifications. Briefly, 3 g of soil were used as an initial sample, and the mechanical lysis was performed with a mixture of glass beads and bead solution that was processed for three pulses of 30 s at 6 m/s in a FastPrep-24 (MP Biomedicals). For both soil types (BS and RZ), DNA from the five corresponding samples was pooled and sequenced using Oxford Nanopore Technologies (ONT). Library preparation was performed using genomic DNA that had been previously fragmented to an N50 of 8,000 bp with a g-TUBE (Covaris), and subsequently processed with the Ligation Sequencing Kit SQK-LSK109 (Oxford Nanopore Technologies) according to the manufacturer's protocol. Sequencing was carried out on a MinION device (version 1B) (Oxford Nanopore Technologies) with an R9.4.1 flow cell. Basecalling was performed using Guppy v6.5.7 (Oxford Nanopore Technologies).