ITS region amplicon sequencing of wheat associated leaf metagenome
The amplicon library construction was done as described previously in Yergeau et al. (2015) based on a dual-indexing strategy that followed the 16S Metagenomic Sequencing Library preparation Illumina guide (Part #15 044 223 Rev. B). For bacterial 16S rRNA gene the V3-V4 hypervariable region was amplified using the universal primers 520F and 799R2 (Edwards et al. 2008), and for fungi the ITS1 region was amplified using ITS1F and 58A2R (Martin and Rygiewicz 2005). PCR products were purified, pooled and submitted for 2 250 bp Illumina MiSeq sequencing at the CESGQ.