Long read ONT 16S Sequencing of Nasal Swab from Chronic Rhinosinusitis Patients
Full length 16S rRNA amplification and sequencing were performed in-house at the Basil Hetzel Institute laboratory(University of Adelaide, Australia). The PCR protocol included initial denaturation 60s(95 degrees Celsius); denaturation 25x20s(95 degrees Celsius); annealing 25x30s(62 degrees Celsius); extension 25x120s(65 degrees Celsius); final extension 300s(65 degrees Celsius); utilising the SQK16S024 Nanopore 16S barcoding kit(Oxford Nanopore, Oxford, United Kingdom). Barcoded amplicons were added into the flow cell for sequencing on the GridION sequencer(Oxford Nanopore, Oxford, United Kingdom). R9.4.1 chemistry MinION flowcells were used. Fastqs were basecalled using Guppy v6.4.6 with super-accurate model configuration.