Validation of PNA clamp for 18S rhizosphere microbiome of maize

We set out to validate a peptide nucleic acid clamp to facilitate the amplification and sequencing of eukaryotic microbes associated with grasses. Here, we tested the ability of the PNA clamp to suppress the amplification of eukarotic microbes present in the maize rhizosphere. We produced libraries using DNA extracted from four maize plants, amplified with or without the clamp for a total of 8 libraries. The libraries were sequenced on the Illumina iSeq 100 platform, with 2 x 150 bp chemistry. The PNA clamp suppressed plant reads from on average 66.5% of total reads to 0.6%, while increasing the number of reads of the microbial animals, protists and fungi.