Invitrogen™ Dynabeads™ In Vitro Transcription and RNA Purification Kit

Features of the Dynabeads In Vitro Transcription and RNA Purification Kit include:

• Flexibility – choose DNA template for IVT with flexibility of design and amplification
• Efficiency – reuse DNA template and beads (at least six times) to help reduce scost and eliminate/minimize plasmid prep hands-on time and risk of bacterial host DNA transfer
• Scalability – swap between manual and automated workflows to match required scale (micrograms to grams of mRNA) and throughput
• Quality – help ensure a safe and flexible supply of magnetic beads to navigate from RNA-based drug discovery research to commercial manufacturing with counterpart solutions (available via the OEM commercial supply), without incurring extra time or costs from switching technology or raw material

Overview of the workflow
A biotinylated template is made by PCR amplification of the target sequence in a plasmid construct or in a synthetic DNA construct using a biotinylated forward primer and a non-biotinylated reverse primer (figure below). As an alternative to PCR amplification, a linearized plasmid can be biotinylated by a fill-in reaction of biotin-dUTP in the 5' overhang sequence, provided the correct plasmid design. The biotinylated template is then immobilized directly to the included Dynabeads Streptavidin for In Vitro Transcription beads without the need for prior purification. The bead-bound template is used directly in the IVT, then removed from the synthesized mRNA by magnetic separation. The template can be reused (at least six times) in consecutive IVT reactions (figure below).

Solid-phase IVT is compatible with commonly used reagents such as the MEGAscript T7 Transcription Kit, mMESSAGE mMACHINE T7 Transcription Kit, and mMESSAGE mMACHINE T7 ULTRA Transcription Kit, based on your needs and strategies for capping and polyadenylation.

Crude RNA from the IVT step is mixed with the included Dynabeads Carboxylic Acid for RNA Purification and the included Dynabeads RNA Binding Buffer is added. The RNA binds to the bead surface and the remaining components of the reaction mixture are removed by applying a magnet (see thermofisher.com/magnets) and discarding the supernatant (figure below).

Automation-ready for use with KingFisher purification systems
The 1-μm magnetic beads are ideal for high-throughput screening. The process can be automated using liquid handlers or any of our KingFisher sample purification systems. Automation with a KingFisher instrument enables IVT template immobilization, IVT reaction, and purification of 96 samples in less than four hours, and up to 5 mg mRNA yield per well. The scripts for KingFisher Flex and KingFisher Apex automated purification workflows are available from our website.

Commercial supply
To learn more about counterpart products and support documentation available for GMP-compliant manufacturing of mRNA vaccines and therapeutics, visit: thermofisher.com/dynabeadsmrna.