mRNA Extraction and Enrichment

Messenger RNA (mRNA) extraction and enrichment can be simplified using magnetic bead-based technology. The method and type of kit used can vary depending on your research needs. For one experiment you may need to quickly obtain high quantities of mRNA, while another experiment may demand the purest quality of mRNA. See below for our selection guide or learn more about other RNA extraction products.

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The selection guide below is designed to help compare kits for different mRNA research applications.

• Method 1: Using oligo (dT) beads: One commonly used technique is poly(A) mRNA selection, which involves selectively isolating mRNA molecules based on the presence of a poly(A) tail at their 3' end. This method utilizes oligo (dT) beads, which bind to the poly(A) tails, allowing for the purification of mRNA from a mixture of RNA molecules. 
• Method 2: Using mRNA capture beads: Another approach is the use of mRNA capture beads, which employ specific probes to target and bind to mRNA molecules of interest. This method enables the isolation of mRNA based on sequence specificity. Additionally, there are commercially available kits that utilize magnetic beads or columns to facilitate the extraction of mRNA. These kits provide a streamlined and efficient process for mRNA extraction.

mRNA has become a powerful tool in fields such as vaccine development, gene therapy, and cancer immunotherapy. One research area utilizing mRNA is in the development of mRNA-based vaccines. These vaccines, such as the COVID-19 vaccines, utilize mRNA to provide instructions to cells on how to produce a harmless piece of the virus, triggering an immune response. Another significant application of mRNA is in the field of gene therapy. Researchers are exploring the use of mRNA to deliver genetic material into cells, potentially correcting genetic disorders or enhancing therapeutic effects. Additionally, mRNA is being investigated for its potential in cancer immunotherapy, where it can be used to stimulate the immune system to target and destroy cancer cells. Overall, the importance of mRNA in these research applications holds great promise for helping advance healthcare and disease treatment.

Oligo(dT)/magnetic capture methods for mRNA purification

Column methods for mRNA purification are reliable but require extensive manual interaction, which means more time and reduced throughput. Magnetic beads conjugated with oligo(dT) offer additional handling and scaling benefits; however, this procedure does require the use of magnetic stands.

The Poly(A)Purist MAG Kit employs the same hybridization and wash solutions as those in the Poly(A)Purist Kit but with the added ease, speed, and scalability of magnetic beads. The protocol typically takes only 45 minutes, and the kit performs from 8 large enrichment reactions (up to 1 mg total RNA input) to 80 small enrichment reactions (as little as 30 µg total RNA input). Like the Poly(A)Purist Kit, this kit only enriches mRNA from previously purified total RNA.

The Dynabeads mRNA Purification Kit for mRNA Purification from Total RNA Preps, enriches mRNA from purified total RNA, typically in 15 minutes, and allows users the flexibility to elute the mRNA from the beads in as little as 5 µL, or even to perform cDNA synthesis directly on the beads without elution.

For those who prefer using Dynabeads technology with sample lysis and enrichment in one kit, the Invitrogen Dynabeads mRNA DIRECT Kits enable 15-minute lysis and enrichment processing of a variety of sample types. In fact, it is possible to capture mRNA and generate cDNA libraries from a single cell using this technology. The flexible format allows users to scale up or down and gives the option to elute the mRNA from the beads in as little as 5 µL, or to perform cDNA synthesis directly on the beads without elution. The 5 mL size is sufficient for 20 standard isolations, while the 10 mL size is sufficient for 40 standard isolations.

For those who work with smaller inputs, the Invitrogen Dynabeads mRNA DIRECT Micro Kit provides enough reagents for 100 mRNA isolations from up to 2.5 x 10⁴ mononuclear cells, up to 1 x 10⁴ cultured cells, or up to 5 mg tissue (depending on the tissue), with the same elution flexibility as the other Dynabeads kits.

Learn more about how you can isolate mRNA in 15 minutes

Bacteria lack the relatively stable poly(A) tails found on eukaryotic mRNA. Until very recently, isolating mRNA from bacteria has been virtually impossible. The Invitrogen MICROBExpress Bacterial mRNA Isolation Kit employs a novel technology to remove >95% of the 16S and 23S rRNA from total RNA of E. coli and other bacterial species. The kit is suitable for mRNA enrichment from ≤10 µg of purified bacterial total RNA in about two hours from a broad spectrum of gram-positive and gram-negative bacteria. mRNA isolated with the MICROBExpress Kit is an exceptional template for synthesizing labeled cDNA for array analysis and is ideal for quantitative RT-PCR, northern blotting, and cDNA library construction.

The most distinctive mRNA purification method takes advantage of specially treated plates in conjunction with optimized reagents to both lyse and enrich mRNA from cells, tissue, blood, or purified total RNA. The Invitrogen mRNA Catcher PLUS product utilizes a proprietary surface treatment of a 96-well plate to facilitate easy and automation-friendly mRNA purification. Locked Nucleic Acid (LNA) technology incorporated into the 20-mer oligo(dT) increases specificity and hybridization efficiency while removing centrifugation, precipitation, and phase-separation steps from the overall workflow.

The mRNA Catcher PLUS Kit is capable of handling 100 ng to 100 µg total RNA, 100 to 10⁶ mammalian cells, up to 4 mg tissue, and 40 µL whole blood. The binding capacity is >80 ng/well. The protocol employs a simple lysis, hybridization, wash, and elution process that can be completely automated on open-format liquid handlers in approximately 1.5 hours from start to finish. The system also has the flexibility to elute mRNA or to perform cDNA synthesis directly in the hybridization plate without elution.