COI Gene Amplification and Sequencing

A 680 bp fragment of the COI gene was amplified using universal primers Fish F1 and Fish R1 and PCR conditions as previously described by Hubert et al. [29 (link)]. PCR was performed with a Phusion® High-Fidelity PCR Master Mix (ThermoFisher, 1040–2678) using the following program: 3 minutes at 98°C followed by 30 cycles of 80 seconds at 98°C, 45 seconds at 55°C and one minute at 68°C, with a final extension of 10 minutes at 72°C. PCR products were sequenced using the BigDye Terminator 3.1 kit (Applied Biosystems). Sequencing was performed on an ABI prism 3130 (Applied Biosystems), at IRD Montpellier, (France) using the Fish F1 primer. Sequences are available on Dryad doi:10.5061/dryad.117tn (see S2 Table).