Miseq analysis of Vitis 140Ru: Root NO-inoculated with cacti inoculum and treated             with Drought Stress_4
Eukaryotic 18S ribosomal RNA (rRNA) gene primers were used in a 28                 cycle PCR (5 cycle used on PCR products) using the HotStarTaq Plus Master Mix Kit                 (Qiagen, USA) under the following conditions: 94C for 3 minutes, followed by 28                 cycles of 94C for 30 seconds, 53C for 40 seconds and 72C for 1 minute, after which a                 final elongation step at 72C for 5 minutes was performed. After amplification, PCR                 products are checked in 2% agarose gel to determine the success of amplification and                 the relative intensity of bands. Multiple samples are pooled together (e.g., 100                 samples) in equal proportions based on their molecular weight and DNA                 concentrations. Pooled samples are purified using calibrated Ampure XP beads. Then                 the pooled and purified PCR product is used to prepare illumina DNA library.                 Sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) on a                 MiSeq following the manufacturers guidelines. Sequence data were processed using MR                 DNA analysis pipeline (MR DNA, Shallowater, TX, USA).