soil metagenome, 60 days point, Daulet Asia
Amplicons were processed using the NEBNext UltraTM II End Repair/dA-Tailing Module (NEB). Barcodes (Native Barcoding Kit 96 (SQK-NBD114.96)) were ligated us-ing Blunt/TA Ligase Master Mil (NEB). Libraries with barcodes were purified using KAPA Pure Beads (Roche, Switzerland). Library concentrations were measured using the Quant-iT dsDNA Assay Kit, High Sensitivity (Thermo Fisher Scientific, USA), and samples were mixed equimolarly. The final adapter (Adapter Mix II Expansion (Ox-ford Nanopore Technologies, UK) was ligated with the pooled library using the NEB-Next Quick Ligation Module (NEB). The prepared DNA library (12 l) was mixed with 37.5 l Sequencing Buffer, and for data collection, 25.5 l Loading Beads were loaded into an R10.4.1 flow cell (FLO-MIN106; Oxford Nanopore Technologies) and se-quenced using MinIONTM Mk1B software version 22.12.7 (Oxford Nanopore Tech-nologies).