Cissy Jiang
RNA-Seq is a powerful tool applicable to an enormous range of biological questions and systems. Although advances in next-gen sequencing and data analysis have improved the accessibility of RNA-Seq, library preparation remains a significant bottleneck in terms of both cost and time. The innovative post-library depletion technology offered by the new SEQuoia RiboDepletion Kit enables the depletion of multiple libraries in a single reaction, maximizing depletion efficiency and saving time and resources.
Ribosomal RNAs (rRNAs) are highly abundant, constituting 80–90% of total cellular RNA. In typical RNA-Seq applications, this large fraction is not of interest, taking up unnecessary sequencing space and reducing the assay’s overall sensitivity. Therefore, efficient removal of rRNA is critical to enable cost-effective sequencing of RNA samples.
A post–library preparation strategy to remove cDNA library fragments derived from rRNA reduces the risk of degrading rare or low abundant transcripts by converting RNA into more stable cDNA early in the workflow. In addition, by eliminating rRNA sequences after library preparation, the SEQuoia RiboDepletion Kit avoids the loss of small, rare, and low abundant RNAs — a problem inherent to pre–library rRNA depletion kits.
The SEQuoia RiboDepletion kit is compatible with most available library prep kits. By bundling the SEQuoia RiboDepletion Kit with the SEQuoia Complete Stranded RNA Library Prep Kit, scientists can profile the entire transcriptome with an easy and streamlined workflow, even from low quantity and degraded RNA samples.
Retain Small Transcripts
Small transcripts, including microRNA (miRNA), small nuclear RNA (snRNA), and small nucleolar RNA (snoRNA), are commonly lost when rRNA is depleted before library construction. A significantly greater number of small RNAs are detected using a post–library depletion method (SEQuoia RiboDepletion Kit) than a pre–library depletion method, resulting in a more complex library and richer dataset.
Depletion of rRNA after RNA-Seq library preparation enriches for RNAs of interest and preserves the diversity of the transcriptome. A significantly greater number of sRNAs (<200 bases) is retained using a post–library depletion strategy compared to a pre–library depletion strategy, resulting in a more complex library and richer dataset.
Pool Multiple Libraries
With the SEQuoia RiboDepletion Kit, you can pool multiple libraries into one depletion reaction to save time and money without compromising depletion efficiency.
RNA-Seq libraries can be depleted in a 96-plex pooled reaction without affecting library quality. Similar depletion efficiency is obtained when libraries are depleted individually compared to when they are pooled and depleted in one multiplexed reaction, even if the libraries are constructed using different library preparation kits.
The SEQuoia RiboDepletion Kit enables researchers to obtain sequencing data that is more representative of the whole transcriptome by employing a post–library depletion strategy, which retains fragments derived from small and long RNA biotypes. Additionally, this product offers significant cost savings by enabling the depletion of multiple libraries in a single reaction. The kit is compatible with most RNA library prep kits, including the SEQuoia Complete Stranded RNA Library Prep Kit.