ITS DNA sequecning of the metagenome of the phyllosphere and thatch of turfgrass treated with different levels of nitrogen fertilizer
The sequencing library for fungal communities was prepared by amplifying the ITS2 region of the rRNA gene using the 5.8S-Fun/ITS4-Fun primers. PCR was performed with the KAPA Biosystems PCR kit, and products were cleaned using the ZR-96 Zymoclean Gel DNA Recovery kit. The amplicons were normalized with the Mag-Bind EquiPure Library Normalization kit, pooled in equal volumes, and quantified using a Qubit 3.0 Fluorometer. The final library was diluted to 4 nM before sequencing on the Illumina MiSeq platform using a 2  300 bp paired-end approach.