In principle you don't need qPCR to look at a miRNA. But don't forget miRNAs are almost the size of one of your primers, so standard RT does not work, you have to ligate a linker with a specific and unique sequence that you will use as template for one primer, using the miRNA as template for the other primer... watch out designing the primers, miRNAs can be very similar within a family.

There are publications in which expression of miRNAs have been studied through RTPCR. In these studies, primers have been made from fold back region to see expression of pre-mirna. Off course, you will not see processed miRNA but it will give some idea about transcriptional regulation of specific miRNA. Try this, may be you get results.

We can look at the expression of mature miRNAs using stem-loop RT-qPCR for miRNAs. Certainly there is no way to see it by normal RT-PCR, it will be too difficult to quantify, such a small piece of RNA on the gel. First we make cDNA of the miRNA by using Stem-loop RT primers and then do real-time on it.

Thank you Peitro, Prabodh and Sachin.

Prabodh Can you recommend me some publications where they have used only RT PCR to check expression of miRNA. I am also wondering if reviewer will trust if we submit the paper with only general RT pcr for mirna expression.

I have designed primers for premiRNA for miRNA, the size of premiRNA being around 60 base pairs, my product size is around 50-55 base pairs. I realised that the actual matured miRNA, which is around 22 nucleotide long is a part of my premiRNA product that I have amplified. It means technically I am doing the right thing. But I am still wondering, it is good enough to convince the reviewer. Please do suggest me further. Thank you.