title: Metabarcoding of root fungal community in Achillea millefolium (plant 2)
design: Experiment2: Plants were grown separately in pots in a glasshouse using a sand:soil mixture. The field-collected soil (2018) was air-dried and sieved, and mixed in a 1:3 ratio with an 2:3 mix of coarse sand and fine sand. Plants were automatically watered and harvested after 4 month of growth. Roots were cleaned and subsampled for DNA extraction (CTAB extraction). From each pot, the soil was thoroughly mixed and a subsample taken for DNA extraction using the Macherey Nagel NucleoSpin Soil kit. 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 Macrogen (Australia) for Illumina sequencing. The raw data is demultiplexed and includes the primer sequences. The numbering of the soil samples (library_ID S01, S02 etc) follows that of their associated root samples (R01, R02 etc), i.e. the sample E2_AchMil1_soil contained the soil from the plant individual E2_AchMil1.