MiSeq of leave metagenome of Haloxylon salicornicum
DNA extractions were performed with the FastDNA Spin kit for soil (MP BiomedicalsTM). The concentration was normalize the DNA concentrations to 0.3 ng.L-1 before PCR amplification. ITS86F (5 GTGAATCATCGAATCTTTGAA 3) and ITS4 (5 TCCTCCGCTTATTGATATGC 3) primers were used to amplify the fungal ITS2. Reactions were performed in 20 L and consisted of 10 L of Buffer Master mix containing Thermo Scientific Phusion High-Fidelity DNA Polymerase (ThermoFisher Scientific), 0.5 L of DMSO, 4.5 L of DNAse free water, 3 L of DNA (0.3 ng.L-1), 1 L of forward primer and 1 L of reverse primer tagged with a short nucleotide sequence. The PCR conditions used were identical for the fungal and bacterial amplifications and were performed in a Veriti 96-well Thermal Cycler (ThermoFisher Scientific) under the following conditions: 10 min initial denaturation, 35 cycles each including denaturation at 94C for 10 sec, annealing at 55C for 20 sec, and extension at 72C for 20 sec, with a final polymerization extension at 72C for 7 min. ITS2 region amplification products were purified using Agencourt AMPure XP beads (Beckman Coulter Inc, Fullerton, California, USA) and performed on a magnetic rack with a 1:1 (v:v) ratio of AMPure XP per PCR product.