title: ITS metabarcoding of native forest and avocado orchard soil
design: ITS2 rDNA region was amplified with the fungal primer system reported in Tedersoo et al. (2014). We used the Taq Platinum Multiplex PCR Master Mix (Life Technologies,USA) protocol. All the PCR reactions contained 12 L of Platinum Multiplex PCR Master Mix (2), 1.8 L of primer mixture (2M of each), 7.5 L of ultrapure DNase/RNase-free distilled water and 1.5 L DNA template (7.5 ng/L). Thermal cycling consisted of an initial step at 95 C for 3 min, followed by 30 cycles of denaturation at 95 C for 45 s, annealing at 60 C for 1.30 min, and extension at 72 C for 1.30 min, with a final extension at 72 C for 10 min.