I have purchased a CircLigase II ssDNA Ligase kit and planning my experiments. According to the ligation reaction mixture (shown in attachment) from the Instructions booklet, only 10 pmol of single stranded nucleic acid can be transformed to cyclic form. In contrast, T4 RNA ligase allows umol scale of single stranded RNA cyclization in one reaction. As my experiments require umol scale of single stranded RNA cyclization, is it possible that I increase the amount of ssRNA in a 20 uL reaction mixture from 10 pmol to 1 umol?
In theory, you could try it and see what happens. But I'd advise you to start by following the instructions in the kit. They are optimized for success.
You can easily dilute ssRNA from a micro molar to pico molar concentration.
You might have to adjust to using a different kit or change the parameters of your experiment.
Good luck!
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