title: Eretmochelys imbricata haplotype 8 anonymous sequence
design: We used the 3RAD protocol as described by Bayona-Vaquez et al. to analyze hatchlings from different nests collected at Abrolhos. We used four of the primers designed by Arantes et al. in this research (3061, 64188, 76958 and 856). The PCR reaction was performed with a final volume of 15 L, following the amounts: 200 M of dNTP, 0.5 units of Platinum Taq DNA polymerase (Invitrogen by Life Technologies), 1.5 mM (up to 4 mM, depending on sample quality) MgCl2, 0.5 M of forward and reverse primers, 0.5 mg/mL of Bovine Serum Albumin (BSA) and 2 L of genomic DNA at 30 ng/L with 1X reaction buffer. PCR cycling was performed with an initial denaturation cycle of 95oC for 5 minutes, 35 denaturation cycles of 95oC for 30 seconds, and variable annealing temperatures for 40 seconds (59C, 56C, 58C and 64C for 856, 3061, 64188, and 76958, respectively), extension to 72oC for 1 minute, and a final extension at 72oC for 7 minutes (Arantes et al. 2020b). The effectiveness of PCR was verified by electrophoresis in a 1% agarose gel and the amplified PCR products were purified by precipitation using a polyethylene glycol (PEG) solution with a concentration of 20 mM.  Sequencing was performed using the BigDye Terminator Cycle Sequencing kit (Applied BiosystemsTM) according to the standard protocol. Amplicons were sequenced on the ABI 3130xl DNA sequencer (Applied BiosystemsTM).