Fungal DNA Extraction and Sequencing
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Corresponding Organization :
Other organizations : Chinese Academy of Sciences, Ningbo Entry-Exit Inspection And Quarantine Bureau
Variable analysis
- Primer pairs and PCR amplification procedures following protocols described by Crous et al. (2009)
- Amplification and sequencing of five loci, including the 5.8S nuclear ribosomal RNA gene with the two flanking internal transcribed spacer (ITS), translation elongation factor (EF-1α), calmodulin (CAM), partial RNA polymerase largest subunit (RPB1) and partial RNA polymerase second largest subunit (RPB2) gene regions
- Genomic DNA extracted from fungal mycelia grown on PDA, using a modified CTAB protocol as described in Guo et al. (2000)
- PCR amplification reaction mixture consisting of 12.5 μL 2 × Taq PCR Master Mix (Vazyme Biotech Co., Ltd, Nanjing, China), 1 μL each of 10 μM primers, 1 μL of the undiluted genomic DNA, adjusted to a final volume of 25 μL with distilled deionized water
- Positive control: None mentioned
- Negative control: None mentioned
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