META-ITS-DIB4-5-6-E05-a
ITS PCR was amplified by using primers and PCR protocols already described in Bartoli et al., ISME J. 2018. PCR products were purified by using cleanNGS beads. Nanodrop was used to quantified the PCR products after purification and BioAnalyzer was performed on 12 randomly selected products to check fragment size. A second PCR was performed with the Illumina indexes by using a equimolar concentration of each PCR product. Products from the second PCr were also purified by using cleanNGS beads and NanoDrop and BioAnalyzer were also performed. Purified PCR products were then merged in an equimolar pool that was quantified by RQ-PCR prior MiSeq Sequencing. Before MiSeq sequencing the PCR pool was further purified by using the cleanNGS beads to eliminate the smell fragments.