Illumina MiSeq microbiome of Bagliettoa sp.
Genomic DNA was extracted using the E.Z.N.A. Forensic DNA kit (Omega Bio-Tek), fol-lowing the instructions of the manufacturer. We used the V1V2 hyper-variable region of the bacterial 16S rRNA as barcode to prospect bacterial diversity. DNA amplification was car-ried out using universal primers 27F and 338R [47]. PCR reactions were performed in a total volume of 15 l, containing 7 l of ACCUZYME DNA Polymerase Mix, 2x (Bioline, Sydney, Australia), 1 l of each primer (10 M), 3 l of distilled water and 3 l of template DNA. PCR settings consisted in an initial denaturation at 95C for 5 min; 30 cycles of 94C for 1 min, 54C for 1 min and 72C for 1 min; with a final extension at 72C for 7 min. PCR products were run in 1% agarose gels stained with SYBR Safe DNA Gel Stain (Thermo Fisher Scientific). Then, they were quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and pooled in equimolar concentrations. Sequencing was carried out by the RTSF Genomics Core at Michigan State University (East Lansing, Michigan), using v2 chem-istry and 2  250-bp paired-end reads on c. 1/10 of a MiSeq run (Illumina, USA).