Applications

• Quick and easy isolation of eukaryotic mRNA that is substantially free of contaminating rRNA, for gene expression analysis, ribonuclease protection assays, RT-PCR, Northern blots, or other procedures.
• Simplified generation of high-quality cDNA libraries.

Note: For RNA-Seq applications, we recommend using a Ribo-Zero™ Kit.

Eukaryotic mRNA that is 3´-polyadenylated has traditionally been isolated by elution from immobilized oligo(dT) or oligo(dU) substrates. The mRNA-ONLY™ Eukaryotic mRNA Isolation Kit* provides a simple and effective method for isolating eukaryotic mRNA that is substantially free of ribosomal RNA (rRNA), in less than an hour, without binding to an oligo(dT) substrate.

The mRNA-ONLY Kit includes Epicentre's Terminator™* 5´-Phosphate-Dependent Exonuclease, a processive 5´→3´ exonuclease that digests RNA having a 5´ monophosphate. The enzyme does not digest RNA having a 5´ triphosphate, a 5´ cap (present on most eukaryotic mRNAs), or a 5´-hydroxyl group. These enzymatic properties make Terminator Exonuclease ideal for producing mRNA-enriched samples from eukaryotic total RNA preparations by selectively digesting the rRNAs (which have a 5´ monophosphate). Thus, the mRNA-ONLY Eukaryotic mRNA Isolation Kit can be used to isolate eukaryotic mRNA substantially free of 5.8S, 18S, and 28S rRNA. Transfer RNA (tRNA), and 5S rRNA are not removed by Terminator Exonuclease digestion.

Benefits

• No oligo(dT)-binding step is required, minimizing losses of mRNA due to inefficient binding.
• The mRNA-ONLY Eukaryotic mRNA Isolation Kit can be used to simultaneously prepare mRNA from both prokaryotic and eukaryotic cells.

*Covered by issued and/or pending patents.