Genomic DNA was extracted from fresh mycelia growing on PDA after 3–5 days of growth following the CTAB method described in Watanabe et al. (2010). For amplification of the ITS, TEF1, GAPDH and RPB2 gene fragments, the primer pairs ITS5/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), gpd1/gpd2 (Berbee et al. 1999) and RPB2-5F/RPB2-7cR (Liu et al. 1999) were used, respectively. A total of 25 μL of a PCR reaction mixture containing 21μL of 1.1×Taq PCR Star Mix (TSINGKE, Beijing, China), 2 μL template DNA and 1μL of each primer was prepared and the PCR was performed in an Eppendorf Mastercycler following the protocols described by Woudenberg et al. (2013). Successful products were purified and sequenced by TSINGKE company (Beijing, China). All sequences were assembled with BioEdit (Hall 1999) and deposited at GenBank (https://www.ncbi.nlm.nih.gov/) (Table (Table11).

Alternaria strains and their accession numbers used in the phylogenetic analysis.

Notes: Alternaria strains of the present study are marked in bold. Type strains are marked ‘T’. Representative strains are marked ‘R’.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.