root-associated fungus metagenome
Full-length fungal ITS region is amplified in duplicate using "fusion primers" (PacBio barcodes + specific region). PCR products are verified visually by running on a high-throughput Hamilton Nimbus Select robot using Coastal Genomics Analytical Gels. PCR reactions from the same samples are pooled in one plate, then cleaned-up and normalized. 190 samples are pooled to make one library which is then quantified fluorometrically before sequencing on a PacBio Sequel II. (design from imr.bio/protocols.html)