qRT-PCR and Northern blot - Is miRNA Northern blot necessary? (Sep/14/2007 )

Hello everyone,

I am right now transfecting MCF7 cell lines with various concentrations miRNA of interest. My question is, after transfection, if I isolate total RNA and do a qRT-PCR for specific miRNA, is that enough? Or do I have to supplement my qRT-PCR data with Northern blots (which I understand is a pain to do!)?

Thanks in advance for your replies.

Thanks,
Una

I think that it is necessary to supplement your qRT PCR data with northern blots.

Best of luck

If you use the ABI TaqMan assays, which specifically amplify the mature miRNA, I'd say qRT-PCR alone is fine. Furthermore, TaqMan is supposed to discriminate between closely related miRNAs, which I think would be more difficult to do with NB.