rDNA ITS2 amplicon sequencing of fine-root samples: plot 4, SS, 6 days after watering
Sequencing was performed on the Illumina MiSeq platform using the ITS2 primer mixes described by  Tedersoo et al. (2015). PCR conditions were 5 min 95C, 28 cycles [30 s 95C, 30 s 55C, 60 s 72C] and 10 min 72C. PCR reaction (total 20 l); 10 l NEB Next High Fidelity Master Mix, 0.5 l of each primer(10 pmol), 8 l of ultra pure water, 1 g of template DNA. Sample purification: Agencourt AMPure XP extraction kit (Beckman Coulter Company), ratio of AMPure XP to PCR reaction is adapted to 1. Indexing PCR: 3 min at 95C, 8 cycles [30 s 95C, 30 s 55C, 30 s 72C] and 10 min 72C. Products are pooled equimolar to a final concentration of 4 nM and sequenced using the MiSeq Reagent kit v3 (600 cycles) for paired end sequencing.