Dna extraction, , PCR

Sun, Bing-Bing, He, Shu-Cheng, Yu, Feng-Ming, Li, Cui-Jin-Yi, Yang, Yan-Yan, Wang, Jian-Rui & Zhao, Qi, 2025, A new species of Periconia (Periconiaceae, Pleosporales) from Xizang, China, Phytotaxa 693 (2), pp. 133-145 : 134

DNA extraction, PCR amplification, and sequencing

Methods for DNA extraction and PCR amplification referred to He et al. (2023). The primer pairs used in this study were: ITS: ITS5/ITS4 ( White et al. 1990); LSU: LR0R/LR5 ( Vilgalys & Hester 1990); SSU: NS1/NS4 ( White et al. 1990); and tef1: tef1 -983F/ tef1 -2218R ( Rehner & Buckley 2005). The total volume of the PCR reaction system is 25 μl, containing 2 μl of DNA template, 1 μl of each primer, and 21 μl of 1 × Power Taq PCR MasterMix. The PCR conditions for amplifying ITS, LSU, and SSU gene loci are: initial denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 95°C for 15 s, annealing at 55°C for 15 s, and extension at 72°C for 15 s, and finally extension at 72°C for 10 min. The PCR reaction program of tef1 is initial denaturation at 98°C for 5 min, followed by 35 cycles of denaturation at 98°C for 45 s, annealing at 52°C for 45 s, extension at 72°C for 50 s, and finally extension at 72°C for 10 min. PCR products were detected by 1% agarose gel electrophoresis. The purification and sequencing were completed by Shanghai Sangon Bioengineering Co., Ltd.