Total S. alopecuroides DNA was extracted using a DNA extraction kit (Fast DNA® SPIN Kit, MPBIO, Santa Ana, USA) and DNA amplification experiments were performed using an ABI GeneAmp® Model 9700 PCR instrument. Fungal primers ITS1F (5’-CTTGGTCATTTAGAGGAAGTAA-3’) and ITS2R (5’-GCTGCGTTCTTCATCGATGC-3’) were used to amplify the ITS1 region of rDNA [37 (link)]. The reaction system was 20 μL: 10×Buffer 2 μL, 2.5 mmol/L dNTPs 2 μL, 5 μmol/L 0.8 μL each for forward and reverse primers, TaKaRa rTaq DNA Polymerase 0.2 μL, BSA 0.2 μL, DNA template 10 ng, and ddH2O was made up to 20 μL. PCR procedure: 95 °C for 3 min; 95 °C for 30 s, 55 °C for 30 s, 72 °C for 45 s; 72 °C for 10 min, 10 °C for holding. PCR products were sampled on 2% agar gel electrophoresis in 3 μL for detection.
The purity and concentration of PCR products and DNA integrity, PCR amplification, and Illumina MiSeq sequencing were performed by Majorbio Co., Ltd. (Shanghai, China). All sequencing data were deposited to the NCBI Sequence Read Archive2 under project number PRJNA832900.
The purity and concentration of PCR products and DNA integrity, PCR amplification, and Illumina MiSeq sequencing were performed by Majorbio Co., Ltd. (Shanghai, China). All sequencing data were deposited to the NCBI Sequence Read Archive2 under project number PRJNA832900.
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