Metabarcoding of fish surimi
The mitochondrial gene region 12S rRNA were amplified by universal primers MiFish-U/E as described [2528]. Each 20L PCR reaction contained 10L of 2x PCRBIO HS Taq Mix, 1L of each primer (10M), 7L ddH2O, and 1L of DNA extract. The following cycling conditions were used: 5minutes at 95C (1); 1min at 95C, 30s at 48C, and 45s at 72C (38); 5min at 72C (1). Three PCR replicates were amplified from each sample and then pooled for a single PCR cleanup with the QIAquick 96 PCR purification kit (Qiagen; 60L elution volume). Agarose (2% w/v) gel electrophoresis was used to verify the amplification of samples. PCR products were pooled and quantified using Qubit dsDNA HS Assay before preparation for the library. The library was sequenced on the NovaSeq 6000 (with paired-end 150-bp reads, PE150) following the NovaSeq XP workflow.