Abi prism 3730xl dna analyser

Fungal mycelium grown on the cellophane of PDA was scraped for the extraction of genomic DNA following a modified CTAB method (Doyle and Doyle 1990 (link)). The primers and PCR conditions are listed in Table 2. DNA sequencing was performed using an ABI PRISM 3730XL DNA Analyser with a BigDye Terminater Kit v.3.1 (Invitrogen, USA) at the Shanghai Invitrogen Biological Technology Company Limited (Beijing, China). The DNA sequences, obtained from forward and reverse primers, were combined using SeqMan v. 7.1.0 in the DNASTAR Lasergene Core Suite software (DNASTAR Inc., Madison, WI, USA).

Genomic DNA was extracted from colonies grown on cellophane-covered PDA using a modified CTAB method (Doyle and Doyle 1990 (link)). The primers and PCR conditions are listed in Table 2. DNA sequencing was performed using an ABI PRISM 3730XL DNA Analyser with a BigDye Terminator Kit v.3.1 (Invitrogen, USA) at the Shanghai Invitrogen Biological Technology Company Limited (Beijing, China). The DNA sequences obtained from forward and reverse primers were combined using SeqMan v. 7.1.0 in the DNASTAR Lasergene Core Suite software (DNASTAR Inc., Madison, WI, USA). Reference sequences were selected based on ex-type or ex-epitype sequences available from relevant recently published literature (Rossman et al. 2007 (link), Suetrong et al. 2015 (link), Norphanphoun et al. 2016 (link), Hongsanan et al. 2017 (link), Senanayake et al. 2017 (link), Voglmayr et al. 2017 (link), Yang et al. 2018 (link), Fan et al. 2018a (link), b (link), 2020 (link)) (Table 1). Subsequent alignments for each gene were generated using MAFFT v.7 (Katoh and Standley 2013 (link)) and manually improved where necessary using MEGA v. 6 (Tamura et al. 2013 (link)). Novel sequences generated in the current study were deposited in GenBank (Table 1, Suppl. materials 1–3: Tables S1–S3) and the aligned matrices used for phylogenetic analyses were submitted to TreeBASE (www.treebase.org; accession number: S24893).

Genomic DNA was extracted from mature colonies grown on SDA plates according to the fungal DNA extraction instructions from the manufacturer (OMEGA) (21 (link)). The Calmodulin (CaM) gene was amplified using primer pairs (F, 5’-CCGAGTACAAGGARGCCTTC, R, 5’-CCGATRGAGGTCATRACGTGG) (31 (link)). The PCR conditions were set as follows: an initial denaturation step of 5 min at 94°C followed by 35 cycles of 30 s at 94°C, 50 s at 57°C and 1 min at 72°C, and a final elongation step of 7 min at 72°C. DNA sequencing was performed using an ABI PRISM^® 3730XL DNA Analyser with a BigDyeTerminater Kit v. 3.1 (Invitrogen, USA) from the General Biology Company (Anhui, China). DNA sequencing was submitted to NCBI, and the accession numbers are listed in Table S1.