RiboMinus technology is designed to enrich the whole spectrum of RNA transcripts by selectively depleting ribosomal RNA molecules (rRNA), regardless of their polyadenylation status or the presence of of a 5'-cap structure. The RiboMinus method has been shown to remove the vast majority of the most abundant ribosomal RNA molecules (up to 99.9%) to allow for greater interrogation of less abundant transcripts.

Working with something other than human, mouse, or plant? Contact us with your species and sequence. Simply include your FASTA sequence in *.txt file format for the ribosomal sequences you would like to verify, with the subject line titled “RiboMinus for (your organism here)”, and we'll work with you to find the right probe for your RiboMinus rRNA depletion.

RiboMinus Workflow

RiboMinus products use a novel purification technology that enriches the RNA transcript spectrum by selective depletion of rRNA transcripts from total RNA.

total RNA purified

Step 1

The protocol begins with total RNA purified from culture, tissues, or blood using a method of choice (e.g., PureLink RNA Mini Kit, TRIzol Reagent).

Hybridize with biotin-LNA probe set

Step 2 - Hybridize with biotin-LNA probe set

The total RNA is then hybridized with Locked Nucleic Acid (LNA™) probes specific for abundant ribosomal RNA molecules.

 

Capture RNA/probe complexes

Step 3 - Capture RNA/probe complexes

Next, the unwanted rRNA are separated from the RiboMinus-enriched complexes using RiboMinus Magnetic beads.

Remove rRNA/probe complexes

Step 4 - Remove rRNA/probe complexes

The RiboMinus-enriched RNA sample is then concentrated using ethanol precipitation or a silica spin column step.

 

Concentrate RiboMinus depleted RNA

Step 5 - Concentrate RiboMinus depleted RNA

The enriched RNA fraction is now ready for downstream processing (e.g., microarray, RNA-Seq).

total RNA purified

Step 1

The protocol begins with total RNA purified from culture, tissues, or blood using a method of choice (e.g., PureLink RNA Mini Kit, TRIzol Reagent).

Hybridize with biotin-LNA probe set

Step 2 - Hybridize with biotin-LNA probe set

The total RNA is then hybridized with Locked Nucleic Acid (LNA™) probes specific for abundant ribosomal RNA molecules.

 

Capture RNA/probe complexes

Step 3 - Capture RNA/probe complexes

Next, the unwanted rRNA are separated from the RiboMinus-enriched complexes using RiboMinus Magnetic beads.

Remove rRNA/probe complexes

Step 4 - Remove rRNA/probe complexes

The RiboMinus-enriched RNA sample is then concentrated using ethanol precipitation or a silica spin column step.

 

Concentrate RiboMinus depleted RNA

Step 5 - Concentrate RiboMinus depleted RNA

The enriched RNA fraction is now ready for downstream processing (e.g., microarray, RNA-Seq).

LNA™ (Locked Nucleic Acid)

The structure of the LNA (Locked Nucleic Acid) monomer consists of a ribonucleoside linked between the 2′ oxygen and 4′ carbon atom of the methylene ring (Braasch & Corey, 2001). This configuration locks the sugar backbone resulting in an increase in Tm (melting temperature). Incorporation of 3 LNA monomers into an oligonucleotide does not affect the ability of the oligonucleotide to bind DNA or RNA but increases the stability of the oligonucleotide/RNA complex (McTigue et al., 2004). Oligonucleotides containing LNA are used in hybridization assays requiring high specificity and reproducibility.

LNA™ (Locked Nucleic Acid)

 LNA is registered trademarks of Exiqon A/S, Vedbaek, Denmark.