I am currently working on DLBCL cell lines, and trying to isolate RNA from exosomes of cell lines. I isolated the exosomes from cell lines using the standard protocol of ultracentrifugation and ultrafiltration. I then went on to isolate total RNA using miRVana and RNeasy kits, but both of them give me negative concentrations. Any suggestions or modifications in the protocol is greatly appreciated!
Thanks,
Hi Shena,
I am routinely using the mirNeasy micro Kit from Qiagen to isolate RNA from exosomes with very good results. In my case, I am working with ECFCs , and I have decent yield in terms of RNA from 80 ml of media. Probably you could increase the volume of media from your cells or the incubation time to get higher exosome concentration. Just as a suggestion I found better results starting from fresh media and not frozen one.
Good luck.
Jose
Thank you very much for your suggestion Jose. I shall do the same.
exosomes have very low RNA content, its normal (if they are pure). see couple papers attached on typical protocols
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