Fig. 2
From: Design and application of a novel two-amplicon approach for defining eukaryotic microbiota
In silico and experimental evaluation of amplicon primers. Predicted DNA primer binding a to eukaryote, archaeal, and bacterial genes and b to protozoan and helminth kingdoms. Primers used in previous published 18S rRNA gene sequencing studies and the highest scoring primers designed for this study are shown for three potential amplicon regions. Tests were performed using SILVA TestProbe3.0 against the v128 Ref_Nr99 (18S) and Ref (28S) databases. c PCR amplification of the V4 V5 amplicon from the 18S rRNA gene using V4-1_F and V4-4_R, or 515f and 1119r primers (top), and the transITS amplicon using V9-9_F and V2-rev6_R primers (bottom). Reactions include a no template control, and template genomes from (1–15) T. gondii RH, N. caninum [NcBahia], B. hominis, T. vaginalis SD7, T. muris G1, T. cruzi [DA], L. panamensis [WR470], G. intestinalis [G3M], E. dispar, S. cerevisiae, C. albicans, A. fumigatus, C. elegans, S. mediterranea, and a human foreskin fibroblast cell line (ATCC SCRC-1041)