Genomic DNA was extracted from entire specimens of B. solaris and P. paranygulgus using the DNeasy Blood & Tissue Kit (QIAGEN), following the manufacturer’s instructions. An 862–base pair (bp) region of the plastid rbcL gene, which encodes for the large subunit of the ribulose-1,5-biphosphate carboxylase/oxygenase, was PCR-amplified using Illustra PuReTaq Ready-To-Go PCR beads (GE Healthcare), and the primers and thermocycling conditions listed in table S1. Amplicons were visualized on 1.5% agarose gels stained with GelRed (Biotium) and enzymatically cleaned before sequencing with Illustra ExoProStar S (GE Healthcare). Clean amplicons were sequenced in 10 μl containing 1 μl of BigDye Terminator (BDT) v3.1 (Applied Biosystems), 2 μl of BDT buffer, 0.5 μM amplification primer, and 1 to 2 μl of PCR product. Cleaned sequencing products were run on an Applied Biosystems 3730S 48-capillary DNA analyzer by the Nucleic Acid Protein Service Unit at UBC. Resulting clean trace files were assembled into full sequences in Geneious v9.1.5 (32) and subjected to a BLAST (Basic Local Alignment Search Tool) search on the National Center for Biotechnology Information (NCBI) website ( and an identification request in the Public Record Barcode Database on the BOLD (Barcode Of Life Data) website ( to verify the plastid’s taxonomic origin.

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