title: Miseq analysis of Vitis 140Ru: Root  inoculated with cacti inoculum and NO  treated with Phosphore Deficiency_5
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.