Metabarcoding ITS2 of root associated metagenome
Total DNA from roots was extracted using the innuPREP Plant DNA Kit (Analytik Jena AG, Jena, Germany) according to the manufacturers instructions. Fungal ribosomal internal transcribed spacer (ITS) region 2 was amplified with a Mastercycler Gradient (Eppendorf AG, Hamburg, Germany) using ITS3_KYO2 (Toju et al., 2012) and ITS4 (White et al., 1990) primers. Purified PCR products were quantified using a Qubit dsDNA HS assay Kit in a Qubit 3.0 Fluorometer (Thermo Fischer Scientific, Dreieich, Germany) and pooled at equimolar concentrations for sequencing. Amplicons were sequenced using the dual index paired-end approach (v3, 2  300 bp) for the MiSeq platform (Illumina).