Abstract
For optimal RNA-Seq results, removal of the ribosomal RNA (rRNA) from an RNA sample is required prior to RNA-Seq library preparation. Commonly used poly(A)+ RNA enrichment methods require the use of high-quality, intact total RNA samples. Yet, even after two passes, the level of rRNA contamination in the RNA-Seq library is often too high. Additionally, poly(A)+ enrichment results in the loss of noncoding, non-rRNA sequences that are an important component of the transcriptome. We describe a greatly improved rRNA hybridization-capture process (Ribo-Zero™ technology) that removes 99% of the cytoplasmic (nuclear-encoded) rRNA and, optionally, mitochondrial and chloroplast rRNA, from both intact and partially degraded RNA samples, including RNA isolated from formalin-fixed paraffin-embedded tissue (FFPE RNA). The Ribo-Zero process recovers mRNA as well as large non-coding, non-rRNAs to enable whole transcriptome RNA-Seq and Ribo-Zero treatment does not alter the expression profile of the RNA sample. We have adapted the Ribo-Zero rRNA removal technology for use with RNA samples from many animal, plant and bacterial species.