title: Podarcis siculus diet study
design: The library preparation was performed with an initial PCR clean-up using Agencourt AMPure XP beads (Beckman Coulter, Brea, CA, USA) to remove primer dimer. This was followed by an indexing PCR, using 7 L of 2 Kapa HiFi, 2.8 L of ultra-pure water, 1.4 L of Index primer (P7/P5), and 2.8 L of cleaned PCR product. This step assigned unique 8 bp dual indices to each amplified product for identification. The thermal cycling conditions were as follows: an initial denaturation at 95 C for 3 minutes; 9 cycles of 95 C for 30 seconds, annealing at 55 C for 30 seconds, and extension at 72 C for 30 seconds; and a final extension at 72 C for 5 minutes. A second clean-up was carried out to eliminate residual primer dimers, nucleotides, and enzymes that might interfere with sequencing. Once these steps had been completed, all the purified PCR products were quantified using Epoch, followed by normalisation to 25 nM, and the PCR products were pooled per marker. Each library was quantified individually using qPCR (KAPA Library Quant Kit qPCR Mix; Bio-Rad iCycler) and then diluted to a concentration of 10 nM. The libraries were subsequently pooled in equimolar proportions and sequenced in a MiSeq desktop sequencer (Illumina) using a MiSeq Reagent Kit v.3 (2250bp), aiming for an average output of 25,000 paired-end reads per sample-marker combination.