High throughput amplicon sequencing fungal p450nor: pooled genomic DNA
Amplification of fungal p450nor gene using primer p450nor394F and 1008R, followed by second amplification using p450nor394F and p450nor809R following protocols of Higgins et al. (2016). Pooled amplicons were ligated to illumina sequencing adapters using Bioo Scientific NEXT Seq DNA sequencing kit and typical quality control (Kapa qPCR quantitation prior to pooling libraries and analyses on Agilent 2100 Bioanalyzer). Sequencing on the Illumina MiSeq was performed as usual for metagenomic samples, but was run in only single read mode since p450nor amplicons are too large for overlap with MiSeq v3 sequencing kit. Hence, 1X600 bp reads were achieved, which should be significantly shortened after trimming and QC.