How do I know what concentration of miRNA probe to use for transfecting a cell line?

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Luigi Mari

Hi, what do you mean with miRNA probe? Are you trying overexpression or inhibition of a miRNA?

Lidia Daimiel-Ruiz

Hi Louisa

For the first transfection experiment I carry out a time and dose curve. Usually I use the dose recommended by the manufacturer and the double and the half (or 2/3 of the recommended dose). And with all doses I perform a time curve using 12 h intervals.

I do that for both overexpression (miRNA mimics) and inhibition (anti-miRNA) and I measure miRNA levels by RT-qPCR.

Then I select the time and the dose with the máximum effect on miRNA levels.

Regards,

Lidia

You could also order miRNA mimic and inhibitor labeled for example with Cy3 (Ambion) and check the efficiency of transfection via flow cytometry, ciaoooo