A: mRNAs modified with the incorporation of N1meΨTP and ΨTP have demonstrated the ability to reduce RNA-mediated innate immune activation in vivo.
A: Yes, the modified nucleotides N1meΨTP and ΨTP are included in their respective kits.
A: The precipitate forms due to some kit components being formulated close to their solubility limits in solution. You may be able to salvage the buffer by heating to 37°C for a short period of time. The easiest way to avoid precipitation of reagents is to assemble the reaction at room temperature (22-25°C) and mix the reactions well prior to the 37°C or 42°C transcription incubation.
A: With lower concentrations of DNA template, you can increase RNA yields by increasing the reaction time from the recommended 1 hour up to 2-4 hours, depending on the amount of template used. Increasing the reaction temperature from 37°C to 42°C also improves the yield.
A: The components of the reagents in a standard 20 μl reaction are very close to their solubility limits. If you set up the reaction on ice, some reaction components (such as the buffer and the nucleotides) will precipitate. After components precipitate, warming the reaction tube only partially resolubilizes the reagents. The T7-FlashScribe™ Enzyme Solution is added last, so it can be kept on ice until needed.
