Hi, I think you are asking about the TaqMan micro RNA assays. Basically you have a stem looped primer for reverse transcription so you can then amplify with a forward known miRNA sequence and a universal reverse primer contained in the stem loop.

I think it is well explained in the ThermoFisher datasheet: (page 27 onwards)

https://tools.thermofisher.com/content/sfs/manuals/cms_042167.pdf

Thank you Daniel,

Yes I know about that. And we are currently using the same approach i.e. specific stem looped primers for each miRNA for their cDNA synthesis. But, I came to know through above mentioned publication that with Universal Stem looped Primer we can make cDNA for all miRNA just like polyadenylation approach. Then we can use a single tube of cDNA to quantify several miRNA similar to random hexamer or oligoDT approach for mature mRNA. These Universal stem looped primer have different neucleotide sequence in their loop taken from Rice genome and random octamer sequence which binds to 3 prime sequence of miRNA and act as primer for one step cDNA preparation.

Oh! I actually have experience with that method but it doesnt involve the TaqMan probes. First you purify the small RNA, you use poly(A) polymerase to polyadenilate all of the small RNAs and then you use a universal primer that ends with oligodT tail to reverse transcribe all of the miRNAs, you can then use the forward sequence of the mature miRNA and reverse universal to quantify the amount of each (Figure 1 of attached paper)

Hope that helps!

Thank you Daniel for your feedback. Sure I will try this approach.

Thanks once again