Remove rRNA and so much more with AnyDeplete

What if you could get rid of all those uninformative transcripts, uncover more of the hidden gems hiding in your data, and reduce your sequencing costs at the same time?

Whole transcriptome RNA-Seq studies are typically hampered by the high levels of background rRNA in the starting material. Frequently this background represents more than half of the total RNA present.

  • What if you could eliminate this annoying background and enrich your final sequencing data for only the informative reads you want?
  • What if you could do this for any sample? From any species? All to your custom specifications?
  • What if you could combine depletion of rRNA from multiple species into a single step?

 

With AnyDeplete technology from Tecan all of this is possible. What's more, by removing unwanted transcripts you can actually lower your sequencing costs while improving your experimental results. Removing the clutter makes it easier to identify the true discoveries hiding in your data.

  • AnyDeplete allows efficient rRNA depletion regardless of input amount from as little as a single cell to a microgram of total RNA
  • Unlike other methods, transcript depletion occurs after library construction, meaning there is no manipulation of input RNA and no bias in the final library
  • AnyDeplete is completely customizable to remove any unwanted sequence in your library, providing high quality, cost-effective data from any sample source

How effective is AnyDeplete?

AnyDeplete dramatically eliminates rRNA reads from your library. As seen in the experiment below, by eliminating the uninformative rRNA, AnyDeplete leaves behind only the sequences you want.

Figure 1. AnyDeplete effectively eliminates rRNA reads leaving only the desired reads providing more informative data. After AnyDeplete, unwanted rRNA reads are reduced to ~ 1% of the sequencing reads from libraries generated with 100 ng of UHR or K562 total RNA. (UHR - Universal Human Reference total RNA; K562 - human immortalized myelogenous leukemia cell line total RNA).

 

 

Do you want to target specific transcripts not covered by off-the-shelf kits?

AnyDeplete designs are customizable for any sample type and any species. With over 15 standard designs covering the most common model species plus humans, and over 170 custom designs completed for over 50 different organisms, AnyDeplete has you covered.

Figure 2. List of custom species targeted using Tecan's AnyDeplete probe design. Some of the designs are proprietary and can not be shared.

 

 

Are you working with single cells or other low input samples and worried about multiple manipulation steps on the input RNA?

AnyDeplete rRNA depletion kits work post library construction. This means that AnyDeplete can effectively remove rRNA reads regardless of the concentration of your starting material. Other kits that use up-front depletion prior to library construction require high inputs limiting the samples that can be studied and involve multiple RNA manipulation steps which can introduce bias. Unlike up-front rRNA depletion methods such as Ribo-Zero, AnyDeplete enables much lower RNA input levels (down to 10 pg) and no RNA manipulation while still offering effective elimination of unwanted reads prior to sequencing. AnyDeplete effectively increases the dynamic range that can be studied, reduces sequencing costs and simplifies data analysis even when starting with ultralow inputs. 

 

 

The simplest, most streamlined workflow for optimal rRNA depletion

AnyDeplete technology is seamlessly integrated into Tecan RNA-Seq library preparation workflows and requires no special operator skills or equipment. Removal of abundant sequences using AnyDeplete is simple and effective.

Figure 3. AnyDeplete target depletion occurs after library construction and is integrated directly into the Tecan protocol.

 

 

Real world application - Whole blood transcriptome profiling of fetal blood

Whole blood mRNA-Seq is challenging due to the high abundance of globin transcripts which can represent up to 80% of the transcripts. Starting with fetal blood is even more challenging since fetal globin is typically not targeted by standard globin depletion panels. 

 

"This type of analysis [whole fetal blood mRNA-Seq] using NGS is challenging due to the high levels of uninformative globin reads in fetal blood. However, the use of the Universal Plus mRNA-Seq kit with adult and fetal globin transcript depletion, provided a simple method to eliminate over 90% of the unwanted sequences, enabling us to detect an additional 1200 low expressed genes with potential implications in pre-term birth"

- Dr. Piotr Mieczkowski, Director of the High Throughput Sequencing Facility, University of North Carolina, Chapel Hill

Figure 4. Universal Plus mRNA-Seq was used to contruct libraries starting with 500 ng of fetal cord blood. In the absence of AnyDeplete, 76.5% of the sequencing reads were derived from globin transcripts representing the majority of the sequencing data. Including adult globin AnyDeplete, reduced the number of globin reads by over 33% providing more informative sequencing data. Customizing the AnyDeplete to include depletion of fetal globin reduced the number of globin reads to 10.4% of the total reads resulting in a significant increase in informative data. The additional informative data resulted in the detection of ~1200 additional low expressed genes providing more comprehensive data without the need for additional sequencing.