circRNA isolation

Question

[dbagmerve]

Is there anyone experienced or knowledgeable about circRNA isolation and cDNA synthesis? I am currently working on a series of experiments in this area and have encountered some challenges, particularly in the steps involving RNase R treatment and cDNA synthesis. How can I optimize the RNase R treatment step? Do you have any recommendations regarding factors such as enzyme concentration, incubation time, and temperature? Is any further action required after RNase R treatment? When performing cDNA synthesis, what should I pay attention to?

[biofalconch]

Sir, this is a Wendy's Bioinformatics forum. Jokes aside, I think it would be a great start by using an already publish protocol (just like this one), you can optimize from there.

[dbagmerve]

Thank you soo much for your reply!