In vitro transcription (IVT) can be a complicated reaction. Scaling up this process requires a good understanding of the various reaction parameters that need to be optimised and controlled to give the best result in terms of mRNA yields and quality.
In this presentation, a review of the results from scaling up the IVT reaction from μg to gram scale mRNA yield will be presented. The effect of several variables on the IVT reaction using different size DNA templates was assessed.
The impact of reaction time, volume, temperature, mixing, reagent concentration, IVT buffer composition and single vs. pulsed addition of reagents on the overall mRNA yield, purity, dsRNA content and capping efficiency will be discussed in detail.