Metabarcoding of soil fungal community in plant-free control soil, experiment 1 (replicate 3)
Experiment1: Plants were grown separately in pots in a glasshouse using a sand:soil mixture. The field-collected soil (2017) was air-dried and sieved, and mixed in a 1:1 ratio with an autoclaved mix of potting soil, coarse sand and pumice (1:2:1). Plants were automatically watered and harvested after 4 month of growth. Roots were cleaned and subsampled for DNA extraction (CTAB extraction). The ITS2 region of the rDNA was amplified by PCR using the primers ITS3 (5-CAHCGATGAAGAACGYRG-3) and ITS4 (5-TCCTSCGCTTATTGATATGC-3) (Tedersoo e al., 2014). The primers included 6nt barcodes for multiplexing. The cleaned PCR products were sent to AGRF (Australia) for Illumina sequencing. The raw data is demultiplexed and includes the primer sequences. Soil controls were plant free and sampled after 4 months as well. DNA was extracted using the MN NucleoSpin Soil kit.