MtDNA genotyping and analyses
The mitochondrial cytb gene was sequenced following the PCR
protocols in Hammer et al. (2014), except that samples were amplified
with the following primer pairs: Glu18 TAACCAGGACTAATGRCTTGAA with
Hd.alt GGRTTGTTGGAGCCTGTTTCAT or Hd.Hyps GGGTTGTTGGAGCCSGTTTCGT and
midg.496 GGCGGCTTTTCRGTAGATAA with Eleo.Thr.40 GATTTTAACCTCCTGCGTCCG.
Sequences were edited using Chromas 2.6.5 (Technelysium, Tewantin,
Queensland, Australia) and imported into BioEdit 7.2.5 (Hall 1999).
Sequences were aligned by eye and checked via amino acid coding in MEGA
to test for unexpected frame shift errors or stop codons. Data were
analysed phylogenetically using RAxML as per for the SNP analysis but
using the model GTRGAMMA.