2.4 Mitochondrial gene amplification and sequencing
A fragment of the mitochondrial gene encoding for the Cytochrome oxidase subunit I (COI ) was amplified using primers C1-J-2183 Jerry (Simon et al., 1994) and C1-N-2568 BEN3R (Brady, Gadau, & Ward, 2000). PCR reactions were performed following Poveda-Martínez et al. (2019). PCR products were checked in 1% agarose gels and purified using the QIAquick PCR Purification Kit (Qiagen, Hilden, Germany). Finally, both strands of the amplicons were sequenced using Sanger methodology at Macrogen Inc. (Seoul, South Korea). Additional COI haplotypes detected in Poveda-Martínez et al., (2019) were retrieved from GenBank (MN013440 - MN013554). All sequences were employed in phylogenetic and population genetic analyses. New mtDNA haplotypes were deposited in GenBank under access numbers: MT138921 - MT138931.