2.6 | Amplification and sequencing
The DMC 1 and rps 16 gene was amplified using the primers listed in Table S1 (Petersen & Seberg 2002; Shaw et al., 2005). All PCRs were conducted in a 50-μL reaction volume, with 1.5 U ExTaq polymerase (TaKaRa, Shiga, Japan). The PCR amplification protocols for the DMC 1 and rps 16 gene are presented in Table S1. PCR products were cloned into the pMD19-T vector (TaKaRa). At least 15 random independent clones were selected for sequencing by Shanghai Sangon Biological Engineering and Technology Service Ltd. (Shanghai, China).