16S rRNA-Seq of TX-S2 pond water sample
Environmental DNA was extracted from each water sample using the NucleoMag Plant Kit. The full-length 16S rRNA gene was amplified, followed by a second PCR run for barcode addition using primers from the Nanopore PCR Barcoding Expansion Kit (EXP-PBC001). The libraries were pooled and subjected to adapter attachment and end-preparation using 60 L of the NEBNext Companion Module for ONT Ligation Sequencing. Library purification and final preparation were performed with the Nanopore Ligation Sequencing Kit (SQK-LSK112). The sequencing was conducted on separate Flongle R10.4.1 flow cells using a MinION sequencing device.