Fig. 2

Comparison of Ribo-Zero Gold (Epidemiology) and Ribo-Zero Plus performance for rRNA removal from total RNA extracted from stool samples versus a synthetic RNA pool containing a mix of bacterial and human RNA. A Overview of the Ribo-Zero Gold (Epidemiology) workflow that captures rRNA using biotin labeled anti-sense RNA probes captured by streptavidin magnetic beads for removal of rRNA from the sample. B Percentage of reads mapping to eukaryotic and prokaryotic rRNAs vs. coding sequences following depletion of adult stool samples and RNA standard with Ribo-Zero Gold (Epidemiology). Boxed bar plots on top represent remaining bacterial rRNA (LSU & SSU) in each sample with a dashed line at the 50% mark. C Overview of the Ribo-Zero Plus method where anti-sense DNA oligonucleotides are hybridized to rRNAs in the sample prior to enzymatic digestion of the rRNA:DNA duplexes with RNase H. D Percentage of reads mapping to eukaryotic and prokaryotic rRNAs vs coding sequences following depletion of adult stool samples and RNA standard with Ribo-Zero Plus. Boxed bar plots on top represent remaining bacterial rRNA (LSU & SSU) in each sample with a dashed line at the 50% mark. E) Diagram of the iterative probe design process, starting from raw sequencing data of samples depleted with the standard Ribo-Zero Plus probe set (DP1). The steps shaded in blue refer to the data shown in Supplemental Figure S3A and B