title: Miseq analysis of Vitis 140Ru: Root PRE-inoculated with cacti inoculum and treated             with Drought Stress_2
design: Distribution of Root-Associated Bacterial Communities Along a                 Salt-Marsh Primary Succession Sequencing of PCR amplicons of 16S rDNA was conducted                 with the Illumina MiSeq platform (CBS, Sfax-Tunisia) targeting the V3-V4 hyper                 variable regions using prokaryotic universal (Pro341/Pro805R) primers sets for 2                 300-bp paired-end sequencing (Illumina). PCR amplification was performed in                 triplicate in 25 l reactions with 12.5 l of 2x KAPA HIFI HotStart Ready Mix (KAPA                 Biosystems), 0.2 M of forward and reverse primers, and about of 5 ng/l DNA. Thermal                 cycling consisted of initial denaturation at 95C for 3 min, followed by 30 cycles of                 denaturation at 95C for 30 s, annealing at 55C for 30 s, and elongation at 72C for                 30 s, finally 72C for 5 min. The PCR products were analyzed on 2% agarose gel.                 Illumina library preparations of the amplified and cleaned PCR products were                 performed. The library quality was assessed on the Qubit@ 3.0 Fluorometer (Thermo                 Scientific) and Agilent Bioanalyzer 2100 system.