Plasmid Linearization For mRNA Production

To guide your R&D programs, Seattle Genova with its significant experience offers linearized plasmids for mRNA applications. Circular plasmid DNA is not suitable for the IVT and production of mRNA, while linearized form is much suitable and stable form for IVT and production of mRNA. The production method starts with the production of a plasmid DNA, then linearized to function as a template to transcribe the mRNA. Linearized pDNA is currently the start line of In-Vitro-Transcription procedures to synthesize mRNA. Seattle Genova will assist you with design customization, selection and feasibility studies. Once designs are finalized our cell line, technique, and quality control assay offerings will assist you to guarantee the high-quality plasmids to meet your desired research needs. 

Mechanism

In order to obtain plasmid linearization, the pDNA is handled in suitable conditions with the ideal restriction enzyme. The particular recognition site for a restriction endonuclease is located after the encoded poly(A) tail, if such exists. After the linearization, the restriction digest is terminated through adding suitable reagents e.g., EDTA, sodium acetate, proteinase K and the reaction is incubated with a purpose to deactivate and eliminate the restriction enzyme. Purification of the linearized pDNA template ensues, generally through silica columns or chromatography, prior to its use as an in-vitro transcription template. Other strategies can also be applied along with phenol extraction, accompanied through ethanol precipitation or centrifugal ultrafiltration. The purified sample including the linearized pDNA is re-suspended in a buffer. A small sample of the DNA may be run on an agarose gel to make sure the linearization of the plasmid takes place correctly.

Highlights

Deliverables

Finalized plasmid is delivered dissolved in sterile TE buffer.

A complete work report.