Foliar mycobiome of relict population of Abies nebrodensis
ITS2-5.8S amplicons which were generated using the target-specific primers 5-CGTCGGCAGCGTCAGATGTGTATAAGAGAC-AGGAACGCAGCRAAIIGYGA-3 and 5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-TCCTCCGCTTATTGATATGC-3. Amplicons concentration was measured with a Qubit BR dsDNA assay kit (Thermofisher) and integrity was assessed by checking 20 random samples with the Tape Station 4150 (Agilent) capillary electrophoresis. Amplicons were indexed according to the Illumina Metagenomic protocol using the Nextera XT Index Kit v2 (Illumina). Final library concentrations were measured with the Qubit HS dsDNA assay kit (Thermofisher) and used to prepare one 7 nanomolar equimolar pool. Libraries were sequenced on a MiSeq platform (Illumina) in a 300 paired-end mode.