What is the best way of profiling microRNA for a clinical condition?

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Quan Li

Microarray?

I assuming you have some patients' samples, serum or ec., do you have healthy and disease controls? Isolate RNAs from your samples.

Since you are handling for microRNAs, please be carefull, do not let your microRNAs degraded. I recommend to use Trizol.

While you got your results back, please confirm those candidates with RT-qPCR.

.......

Good luck!

Ajay Saini

I also agree with Quan Li....go for miRNA microarray analysis ....to identify miRNAs in a particular condition...then validate them by qRT-PCR.....I'll also add that ....also validate the down-regulation of the target gene of the miRNAs of interest in the same samples...to get some more insights..

bye

Eva-Maria Sedlmeier

Hi, you can also profile your microRNAs with low density arrays by RT-qPCR. They are also called TaqMan Arrays. But you have also to validate your selected candidates afterwards.

Regards,

Eva

Harsh Dweep

Read below review article for more information:

http://circres.ahajournals.org/content/108/2/219.full.pdf+html

Thomas Litman

I would suggest miRNA microarrays for screening, and then validation by Q-PCR. Depending on how many samples you have, you could consider small RNA-seq as well, by multiplexing (barcoding) your samples. Another important issue is your choice of tissue: Are you working with fresh-frozen or FFPE samples? Pick one, and stay with this, as you may encounter problems when migrating from one to the other. This also applies to the RNA isolation procedure: different extraction methods WILL give very different results ... Finally, if your "clinical condition" (such as a tumor) has a corresponding "normal adjacent tissue" control, include this, as it will give you an idea of each patient's baseline levels.