Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps) (Invitrogen™)

The Dynabeads™ mRNA Purification Kit rapidly isolates the mRNA transcriptome in typically 15 minutes, delivering pure, intact mRNA. The kit is designed to specifically target, capture, and purify mRNA molecules from total RNA preparations. Advantages of using the Dynabeads™ mRNA Purification Kit:

• Obtain pure mRNA in typically 15 minutes
• Recover and enrich the transcriptome efficiently
• Prepare mRNA that is suitable for nearly every downstream application

Pure, rapid mRNA isolation
The Dynabeads™ mRNA Purification Kit contains magnetic beads for the isolation of the mRNA transcriptome from any total eukaryotic RNA preparation (see figure). Dynabeads™ magnetic beads are uniform in size and highly mobile in solution (see figure). This enables the bead capture surface to quickly and continuously interact with the entire total RNA sample during the mRNA capture phase. The ribosomal RNA and small RNA molecules (transfer RNA, microRNA, small nucleolar RNA, and small cytoplasmic RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured resulting in cleaner, more sensitive results (see figure). Within about 15 minutes, pure mRNA is isolated and ready for use in downstream applications.

A straightforward enrichment procedure
The Dynabeads™ mRNA Purification Kit uses a robust affinity purification principle for the enrichment of polyadenylated mRNA. Superparamagnetic Dynabeads™, coupled to oligo-(dT)₂₅, are first equilibrated with Binding Buffer, and then mixed with purified total RNA. The beads are then washed to remove contaminating RNA species, and then mRNA is eluted in as little as 5 μL of 10 mM Tris-HCl. The entire process is facilitated by the use of a neodymium magnet (purchased separately), which allows for the quick and efficient immobilization of the magnetic beads during buffer changes. RNA is suitable for all downstream molecular applications, including:

• Gene cloning
• cDNA synthesis, cDNA library construction
• RT-PCR, quantitative RT-PCR
• RPA (Ribonuclease Protection Assay)
• Subtractive hybridization
• Dot/slot hybridization
• Primer extension