Hello

This is depend on the source of exosomes. Please , visit websites & links here will help you

Good Luck

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0046874

http://www.hindawi.com/journals/dm/2015/893594/

http://www.sabiosciences.com/manuals/Whitepaper_Serumplasma.pdf

http://exosomes.gene-quantification.info/

http://www.hindawi.com/journals/bmri/2015/492395/

http://montgomerylab.stanford.edu/research.html

http://exosomestalk.com/

 Exosomes-enriched proteins include members of the tetraspanin family (CD9, CD63 and CD81), members of the endosomal sorting complexes required for transport (TSG101 and Alix) and heat-shock proteins (Hsp60, Hsp70 and Hsp90) can be the candidates of the internal control of exosome analysis. 

thank you! this very helpful

There is a similar post asking this same question, in which they recommend to use miR-16 and let-7d as controls. In my case, since I want to validate some results from a NGS experiment I took 8 highly expressed miRNAs that are stable (and these two were amongst them). Also, a widely used technique used in many groups and studies is to use >15 housekeeping miRNAs and normalize your miR of interest against the average Ct of all of them.

Hi Jennifer,

I have a question regarding the controls for normalization. Let's suppose that I'd like to validate the NGS results obtained from exosomal RNAs (healthy vs. patients serum). How reliable is the normalization using highly expressed and "stable" miRNAs??  I can understand that using more than 15 miRNAs can minimize the error caused by single miRNA (which actually makes me doubt about their "house-keepingness"), but using this strategy (the more miRNAs for normalization, the better) the costs are getting higher. Isn't there some cheaper option? Do you know some papers about circulating miRNAs that are reported as "stably expressed" in plasma/serum? For me it would be very helpful :) Frankly I think it's a pain in the ass (sorry for the expression) to find a reliable control for qPCR...and not to talk about the diverse results (from the same sample) we can obtain by using different RNA extraction methods...

Hello Renata, in my opinion, if you use serum directly, not purified exosomes, you could use a spike-in before the RNA extraction and use it as "housekeeping".

Despite this, it is never recommended, at least in miRNAs studies, to use a single housekeeping miRNA/smallRNA, for me in cellular PCRs use 2-3 RNUs.

About how reliable it is to use highly stable miRNAs, it will depend on your experiment. I have chosen those that are highly expressed on exosomes and stable in all the three conditions I study. This list I got it from my NGS data itself, if you only have one single miRNA that you think it is stable... how are you going to be sure if you only study this one? I validate the stability of all these miRNAs at diff concs and saw that they were all very stable so... Also I was lucky that my group is specialized in miRNAs and we have a great stock of primers so I could validate them without having to invest any more money.

And I think I have read somewhere that they used some RNUs for normalization, and according to my data they are not stable at all even between replicates, so.... You must decide what do you want to do, but I think that in your case a spike in wouldn't be a bad idea.

Thank you very much for your answer, it was very helpful :) In our laboratory we are still not decided whether to perform RNA profiling from serum or from isolated exosomes and which one would be better for diagnostic purposes....we are just trying to set up the best option for our experimental design, analyzing potential pitfalls....and yes, we usually use 2 controls in PCR, but well... you can never be 100% sure :)