DNA Extraction and Sequencing Workflow

The DNeasy® Blood and Tissue Kit (Qiagen) were used to extract genomic DNA from each individual, following the manufacturer’s instructions. Primers were designed using Primer3 [46 (link)] (Additional file 7). PCR reactions were performed using 100 ng/uL template DNA, MyTaq™ Red Mix (Bioline) and 1 pmol of primers, with initial denaturation at 94°C for 2 minutes, followed by 36 cycles of: 30 seconds at 94°C, 30 seconds at 50°C-65°C, 60 seconds at 72°C; then 72°C for 10 minutes. Resulting products were separated and purified using the PureLink® Kit (Invitrogen). DNA sequencing was performed by the Biomolecular Resource Facilty (The Australian National University), or the Ramaciotti Centre (University of New South Wales). Sequence trace files were visualised using 4 Peaks software from Nucleobytes (www.nucleobytes.com) and sequence alignments were performed in ClustalX 2.1 with default parameters.