Rapid Sequencing gDNA-Barcoding Kit
The library design involved a rapid barcoding-based, single-end metagenomic sequencing approach using the Oxford Nanopore Technologies Rapid Sequencing gDNA-Barcoding Kit (SQK-RBK110.96). Each sample was prepared with 55 ng of template DNA and 1 l of a unique rapid barcode, allowing for multiplexing. Barcoded samples were pooled and purified using AMPure XP Reagent for size selection. After ethanol washing and elution, Rapid Adapter F (RAP F) was added to the pooled DNA. The final sequencing library was prepared by mixing the DNA with Sequencing Buffer II and Loading Beads II, then loaded onto an R9.4.1 MinION flow cell. Sequencing was performed for 12 hours, following the manufacturers protocol for rapid and efficient metagenomic data generation.