A D V E R T I S E M E N T
How to make RNA get linear and prevent its secondary structure?
In order to linearize or denature RNA, it can be achieved by heating to 80-90o for 5 minutes, then cooling quickly on ice (taking care not to cause precipitation of the loading-buffer components) and immediately applying it.
The step above can reduce the RNA secondary structure to be formed and also aid binding of the poly (A+)-tailed mRNA to the oligo (dT).