16S rRNA gene nanopore seq bulk soil: location 2; biocrust 2
PCR amplification of the full-length bacterial 16S rRNA gene (V1-V9; ~1.45 kbp) was carried out by using the S-D-Bact-0008-a-S-16 and S-D-Bact-1492-a-A-16. Barcodes were added by employing the PCR Barcoding Expansion Pack 1-96. The Ligation Sequencing Kit (ONT, Oxford, UK, Cat. No.: SQK-LSK109) was employed to prepare the sequencing library. A R9.4.1 MinION flow cell (ONT, Oxford, UK, Cat. No.: FLO-MIN106D) was primed and loaded as indicated by the manufacturer. Sequencing was performed during ~6.5 h. Reads were basecalled with MinKNOW software (v. 20.06.5; core v. 4.0.5) using Guppys (v. 4.0.9) fast basecalling model, and sequences with Q < 7 were discarded.