I'm evaluating microRNAs expression in neuronal cells-derived exosomes. I was wondering which microRNA I should use as control to normalize my results.
Thanks in advance
Hi Patricia
Not too many choices for doing that. Spike-in with a synthetic and unrelated miRNA will be a good solution.
However, if you know some miRNAs in your exosomes that are not changing in your particular process, you can use them as well.
Good luck. Best
Paco
One can get free protein bounded rna/miRNA in the exosomes preparation.If you use any RNA/miRNA as internal control...One need to show if that RNA is protected ....i guess thats needed to show if we are normalizing with correct RNA ..
This could be one reason that we don't have proper internal control...In my knowledge No methodological paper on exo rna normalization :(
Or Absolute quantification can be used..
Instead of normalizing I suggest you try absolute quantitation using miRNA standard curves. IDT can synthesize the miRNAs for you using the sequences from miRBase or if you want a more stable standard, Dharmacon's miRNA mimics could be used as standards also. If determine your total RNA concentration (assuming you haven't using a purification method that isolates a miRNA enriched fraction which cannot be accurately quantitated using a spectrophotometer such as nanodrop) and use equal amounts of starting material for qPCR you should be able to accurately quantitate the miRNAs present in your sample in a reproducible manner.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Nucleic Acids
- RNA
- Small RNA
- microRNA