Optimizing conditions for efficient long RNA synthesis
Several factors are very important to the good working of the synthesis of RNA. Most notably, the quantity of magnesium ions in the buffer itself is important. RNA polymerase requires a sizable amount of Magnesium to support proper interaction with the DNA template and subsequent folding of the RNA. A second important aspect to consider is the concentration of DNA template. Specifically, you must have enough template DNA reaction in order to get good RNA yields.
Long RNA, of course, will come with its own set of issues. That these problems can relate to the breakdown of the RNA, affect how the RNA folds, and build-up structures that potentially interfere with synthesizing the RNA. One way to address these challenges is by tweaking the buffer you take. For a more comprehensive structure of RNA, researchers can add different special chemicals such as DTT and p-mercaptoethanol to help the RNA fold correctly and prevent it from sticking together, or clumping.
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