LR-PCR of Euchloe ausonides subsp.
LR-PCR primers were adapted from Park et al. (2012) to better match Euchloe sequences in GenBank. Each mitogenomic fragment was amplified in a 50  PCR reaction consisting of 0.5  TaKaRa LA Taq (5U/ stock concentration), 5  10X LA PCR Buffer, 5  25 nM MgCl2, 8  dNTP mixture (2.5 mM each stock concentration), 1  forward and 1  reverse primers (each at 10  stock concentration, and 0.4  final concentration) 2  DNA template, and 27.5  PCR grade water. Thermal cycling parameters for each of the fragments were 94 for 1 min, followed by 35 cycles of 98 for 10s, relevant AT from Table S3 for 15 min, 72 for 30s, and a final extension at 72 for 10 minutes. These fragments were purified and pooled in equimolar concentrations and sequenced on the MiSeq.