Overview of the Fungal Metagenomic Sequencing Demonstrated Protocol
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The Fungal Metagenomic Sequencing Demonstrated Protocol is for the analysis of fungal metagenomic samples, and describes primer sequences and an assay workflow for use with a BaseSpace Sequence Hub (BSSH) analysis solution. The protocol details a 2-step PCR amplicon workflow for the amplification and analysis of the Internal Transcribed Spacer 1 (ITS1) region. The current BSSH analysis solution is 16S Metagenomics v.1.1.3, which includes the UNITE Fungal ITS Database v7.2 reference taxonomy database.
Note: If using the recommended primers or other sets of primers, dilute an aliquot of each primer to 1 µM with 1X TE pH 8 buffer or nuclease-free water. Pool equal volumes of each primer to create a forward primer pool and a reverse primer pool. Aliquot 5 µl of each primer pool into each library plate well containing sample. There are 8 forward and 7 reverse primers in the Illumina design.
Resources:
Uses indexes from the sets A - D (Illumina, Catalog No. FC-131-2001, FC-131-2002, FC-131-2003, or FC-131-2004)
BaseSpace Demo Data for ITS from App Note:
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