85 Table 4 SI Appendix. Primer sequences, associated dye and PCR condition for the nine microsatellite loci used to assess connectivity between the RP and GLS loggerhead turtle nesting areas. ¤ = position of the corresponding dye. PCR = letters refer to PCR thermocycler conditions summarized in Table 5 SI Appendix. Locus Forward Primer (5’ to 3’) Reverse Primer (5’ to 3’) Dye PCR Cc1F01 ¤GTGTGAAGGCTCTAAAACTAAT GTTTATACTGGGACGATAGGATAAA 6-FAM C Cc5F01 GTTTAAAGGATTTGAGATGTTGTATG ¤CCAGTTGTCTTTCTCCAGT VIC D Cc7B07 GTTTATAATGTTGGTGAGCAATATAG ¤GGAGTTAAACCAGGCACAGT VIC A Cc2G10 ¤GTGGCAAGGTCAAATACAG GTTTGCCCTTATTTGGTCACA PET B Cc117 ¤TCTTTAACGTATCTCCTGTAGCTC CAGTAGTGTCAGTTCATTGTTTCA 6-FAM A Cm84 GTTTTGACATTAGTCCAGGATTG ¤ATTGTTATAGCCTATTGTTCAGGA VIC A Cm72 ¤CTATAAGGAGAAAGCGTTAAGACA CCAAATTAGGATTACACAGCCAAC NED A Cc141 ¤CAGCAGGCTGTCAGTTCTCCAC TAGTACGTCTGGCCTGACTTT NED A Cc7 ¤TGCATTGCTTGACCAATTAGTGAG ACATGTATAGTTGAGGAGCAAGTG 6-FAM A One primer for each pair was fluorescently labeled with NED, PET, VIC or 6-FAM. Each Polymerase Chain Reaction (PCR) had a final volume of 15 µL and included 1 X AmpliTaq GoldTM 360 Master Mix, 0.4 µM of unlabeled primer, 0.5 µM of labeled primer, and 1-10 ng of template DNA. Four conditions were performed to amplify loci (A to D). Table 5 SI Appendix. The four PCR thermocycler conditions used for the genotyping of the nine microsatellite loci. Indicated letters refer to target loci listed in Table 4 SI Appendix.
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