DNeasy blood and tissue kits were used to extract DNA from
fin tissue by following the manufacturer’s instructions (Qiagen).
Ten gene regions in both the mitochondrial genome (mtDNA) and
nuclear genome (nuDNA) were targeted for DNA barcoding. MtDNA
gene regions included CO1, the control region (CR), cytochrome b
(Cytb), ATP synthase (ATPase), and NADH dehydrogenase 2 (ND2)
and 4 (ND4). NuDNA gene regions included recombination activating
genes 1 (RAG1) and 2 (RAG2), histone protein 3a (H3a), and ring
finger protein 213 (RNF213). The choice of these markers was based
on a combination of previous experience with the genetic identification
of inshore fisheries species in Queensland, the availability of
universal primers, and prior success of PCR amplifications in other
Sillago species.