Miseq  of Opuntia ficus-indica: Roots isolated from spineless plant of Bizerte
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.