In this study, two different FFPE RNA samples were treated with either the Ribo-Zero Kit (Human/Mouse/Rat), which removes cytoplasmic rRNA, or the Ribo-Zero Gold Kit (Human/Mouse/Rat), which removes both cytoplasmic and mitochondrial rRNA. The rRNA-depleted RNA was then used to prepare RNA-Seq libraries using the ScriptSeq v2 Kit. Libraries were sequenced on an Illumina® GAIIx sequencer.
|Figure 1 (click to enlarge). Summary of sequencing metrics from FFPE RNA libraries.|
As shown in Figure 1, the workflow yields high-quality RNA-Seq libraries, with excellent removal of rRNA and >98% directionality from FFPE RNA. Quality metrics surpass those obtained from FFPE RNA using standard ligation-based library preparation workflows.