Nanopore sequencing of quantitative PCR assays reveals off target Aspergillus species commonly found in Cannabis flowers

Illumina Sequecing of a qPCR assay was performed. Paired Reads were merged with FLASH, trimmed and q30 quality filtered and BLAST'd against NCBI NR. Krona plots were generated with the top 1000 reads. <br> For ONT analysis, 2 different ITS amplicons (450bp and 750bp) were amplified from the Pre-PCR DNA to assess the microbiome diversity the qPCR assay was targeting. <br> The 450bp ITS assay is fungal specific. The 750bp ITS assay include plants and fungi (Cheng et al). <br> Amplicons were converted to Barcode ready sequencing libraries and sequenced on a Flongle with SUP basecalling mode. Reads &gt;Q14 and 200-800bp in length were BLAST'd and fed into Krona plots.