Metagenome of Soil
Extracted DNA samples were amplified using the full length of 16S rRNA gene primers with common primer binding sequences 27f and 1492r, attached to unique 24 bp barcodes and nanopore motor protein tether sequence. The PCR was performed with 600 nM of each forward and reverse primer, 25 L of Premix Taq DNA Polymerase, and a 10 L DNA template in a 50 L reaction. The amplicons from the PCR step were purified using NucleoSpin Gel and PCR Clean-up. The barcoded amplicon samples were pooled in equimolar ratios, and library preparation and sequencing were conducted using Ligation Sequencing Kit SQK-LSK-109