18s amplicons covering V4 and V9 hypervariable regions from Anopheles colutzii samples from West Africa (Larval, Unfed adults, Human Bloodfed, Animal Bloodfed) amplified with two PCR clamp approaches to maximize amplification of eukaryotic microbes.. Preferential suppression of Anopheles gambiae host sequences for mosquito eukaryotic microbiome profiling

The identification of the eukaryotic microbiota by targeting the 18s rRNA gene is challenging due to simultaneous amplification of the abundant 18s rRNA gene target in the eukaryotic mosquito host. Here, we use defined panels of A. gambiae samples from West Africa to test two experimental PCR clamp approaches to maximize the specific amplification of 18s rRNA gene hypervariable regions from eukaryotic microbes using: i) anneal-inhibiting blocking primers (annealing blockers), and ii) peptide-nucleic acid (PNA) oligonucleotides (PNA blockers).