I am planning to get some blood from patients to perform RNA seq (circular RNA which requires deep sequencing). I am wondering if anyone can share the protocol with details. I am also wondering about the amount of blood (whole / plasma / serum) to isolate RNA for deep sequencing. Thank you so much for your help!
i'm guessing you are looking at circulating RNAs? not PBMCs? I have heard that 1ml of plasma/serum is good for exosomes, but process it well to remove all RBCs - spin and transfer 90% to a new tube 2-4 times to ensure no RBCs in the mix. yes, you will leave some behind but cleaner is better than just more. so if you need 1ml of plasma/serum then you would need 2-3ml of whole blood. check the exosome literature for some protocols
You can use EasyPure® Genomic DNA Kit, 200ul from clotted blood is enough to start your protocol then follow the pamphlet protocol
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