My cRNA yields are consistently low even though my total RNA quality and cleanliness appears to be very good. What is the problem?
Likely when you are setting up your IVT reaction you are not letting the components warm up to room temperature. You should let ALL the components of the reaction (including cDNA and with the exception of the enzyme) warm up to room temperature. The reaction buffer contains spermidine, which can precipitate cDNA in cold solutions.
