Development of Illumina oligos for fungal ITS sequencing

New PCR primers were designed with the following objectives: 1) targetting the ITS2 region to minimize bias due to introns at the 3' end of the SSU, 2) have the broadest possible coverage of Fungi, 3) minimize amplification of plant, protist and other Eukaryote sequences, 4) compatibility with Illumina sequencing adaptors and GOLAY barcodes. Primer regions were selected by eye from sequence alignments then further analyzed using PrimerProspector. At present, several mock communities and hundreds of soil DNA extracts have been successfully sequenced on the Illumina MiSeq platform using the oligos that were developed.