Hi,
Can anyone suggest good endogenous control RNU? for serum based exosomal miRNA ? I used RNU48, 4427975 TaqMan from thermofischer which did not work cT value above 35.
Hello,
There isn't a universal normalizer for serum derived exosomal miRs to be honest. It may depend upon your sample. But a lot of people use miR16 which again varies itself a great deal. Qiagen recommends C. elegans miR-39 as a spike in control, which is at least found better than others in many cases but again this is not an absolute statement.
Thanks
Hi there
I strongly recommend not to use natural normalizers. They are extremely variable and dependent on the circumstances. Use any spike-in synthetic RNA as Rajeev suggested.
In the particular case that you are doing a screening of miRNAs and you quantify more than 50 you always can use the "global normalization" approach.
Best. Paco
As suggested above, I also favor the use of spike-in instead of taking any miRNA as a normalization control. RNU6B or similar are more suitable when origin of miRNAs is other (tissue etc) than body fluids as described in our recent article, http://www.nature.com/articles/srep21510
Best, Azeem
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Nucleic Acids
- RNA
- Small RNA
- microRNA