Coverage across the T. rathkei mitochondrial genome was quite high, >300× across most of the genome in each sample, except at the very ends of the monomer sequence (Figure 2). There were no fixed sequence differences between the two individuals sequenced. However, eight heteroplasmic sites were identified (Figure 2, Table 1). Six of these heteroplasmic sites were detected with confidence in both individuals, and each allele occurred at a frequency of approximately 0.5 in all of these cases. Three of the six occurred toward the ends of the assembled monomer sequence (positions 62, 73, and 14107), and thus had relatively low coverage. The remaining three occurred in putative tRNA genes, and in the anticodon of each (positions 9346, 11785, and 12188). These three were supported by high coverage in both individuals; we also confirmed all three in additional unrelated wild-caught individuals using Sanger dye-terminator sequencing (Figure S1).