18 S metabarcoding of Trypanosoma: animal sample
Genomic DNA was extracted from host tissues and blood using commercial purification kits. A target fragment of the 18S rRNA gene was amplified via a nested touchdown PCR approach, utilizing primers SSU4_F and 18Sq1R for the initial amplification round, followed by primers SSU561F and SSU561R for the second round. The resulting amplicons were barcoded and multiplexed employing the Rapid Barcoding Sequencing Kit 96 V14 (SQK-RBK114.96) following the manufacturer's protocol. Sequencing was executed on an Oxford Nanopore Technologies MinION Mk1C platform equipped with an R10.4.1 Flongle Flow Cell. Raw ionic current signal data was processed, basecalled, and demultiplexed using Dorado with the High-accuracy model.
SUB16272240
90