Yes, we would be happy to use your DNA plasmids for the synthesis of your mRNA sequence. For us to utilize your plasmid in the synthesis of high-quality mRNA, your plasmid should meet the following requirements:
• T7 promoter upstream of the 5′ UTR
• T7 promoter should preferably be followed by three G nucleotides ….TATA(GGG)
• A 5′ and 3′ UTR sequence
• Linearization site after 3’ UTR. Preferably a type IIS restriction site creating a 5’ overhang.
• Kozak site (GCCACC(ATG)

If you are unsure whether your plasmid meets the requirements, feel free to contact one of our Product Specialists, they are happy to help!

At -20°C the mRNA is stable for approximately one month, while at –80°C the mRNA is stable for at least a year. We advise to aliquot the mRNA sample the first time you thaw the mRNA, so that the efficacy and stability of the product does not decrease by repeated freeze-thaw cycles.

We can synthesize custom mRNA sequences starting from 100nt up to 15k nt.

After the formulation and dialysis we measure the size distribution of the LNP sample via Dynamic Light Scattering (DLS), we also have the option to measure the zèta potential (electrokinetic potential) and encapsulation efficiency assay upon request. An internal control LNP is always formulated during a production run and allows us to validate the activity and toxicity of the LNPs on HeLa cells.

We measure the purity of mRNA by spectrophotometry (A260/280>2, A260/230>2.2). We also validate the size of the mRNA product and the absence of any by-products via RNA gel electrophoresis. An internal control mRNA is always synthesized during a production run and allows us to validate the activity of the sample on HeLa cells.

As the dose is cell-type-dependent, the dose you use for one cell-type might not give you equal results in another. Therefore, we advise to test and try at least 3 doses of mRNA and determine linearity for the specific cell type you are using. A safe range to stay in for a well-designed mRNA is around 50-70 ng/96-well, although some cells will tolerate up to 200-250 ng per 96-well. At RIBOPRO, we often use 10, 50, and 100 ng to determine the optimal dose. Keep in mind that the dose scales (linearly) with a larger surface of cells.
For in vivo use, the species and the target organ make a huge difference in the recommended dosing. For mice, we generally recommend staying below 1-2ug/25gram animal.