Genjuice_2021_Tele02_12S
Extracted DNA was PCR amplified targeting a ~167 bp fragment of the mitochondrial 12S gene using the fish-specific Tele02 primers (Taberlet et al., 2018). PCR products were then purified with Mag-Bind  TotalPure NGS magnetic beads (Omega Bio-tek Inc), adding to 30 l of PCR products a 1 ratio of magnetic beads (Bronner et al., 2009). Purified DNA was quantified using a Qubit 4.0 fluorometer with the Qubit dsDNA HS Assay Kit (Invitrogen). Based on the total DNA concentration, samples were normalized and pooled in equimolar concentration for library preparation. End repair, adapter ligation and library PCR amplification were performed using the NEXTFLEX Rapid DNA-Seq Kit 2.0 for Illumina platforms (PerkinElmer) according to the manufacturer's protocol. The Agilent 2200 TapeStation and High Sensitivity D1000 ScreenTape (Agilent Technologies) indicated secondary products (e.g., adaptor dimers) remained, which were removed by another magnetic bead clean-up (1x ratio). The library was quantified using quantitative PCR (qPCR) on a Rotor-GeneQ (Qiagen) with the NEBNext Library Quant Kit for Illumina (New England Biolabs) and then diluted to 1 nM. The final library was sequenced at 60 pM with 10% PhiX Control on an Illumina iSeq 100 using the i1 Reagent v2 (300-cycle) (Illumina Inc.).