High throughput rRNA sequencing of free-living amoebae and their associated microorganisms
DNA extracts were subjected to high fidelity PCR using the Q5 DNA polymerase. Amplicons were purified using a silica membrane column-based kit. Amplicons were quantified, adjusted to 200 fmol each and processed for native barcoding kit (SQK-NBD114.96) according to the manufacturer's instructions (protocol version NBA_9170_v114_revE_15Sep2022). Sequencing was done on a MK1C MinIon, in high accuracy mode.