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).