I would like to look at the expression of a single miRNA. At this point, I would be inclined to look at miRNA expression by general RT (similar to RT for any other mRNA for a gene) without going through qRT PCR. would it be possible to do that? Are there any publication/s which has done the same? I would be grateful if anybody could suggest me. Thanks.
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.
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