Hi Richard

You you'are able to get hold of the initial papers on the miRNA quantification usinf different approaches...you'll get the details about all aspects of the reaction.....basically they only differ in the ways developed to detetc very small RNA molecules...rest everything is similar to a regular qPCR..

bye

So as far as i can tell you have to pay a biotec company to prepare the inital probes that extend the microRNA's and then you perform QRT-PCR on those extended fragment. Is that right? For example here are a couple of companies offering the service...

http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/epigenetics-noncoding-rna-research/miRNA-Profiling-/miRNA_qRT_PCR/TaqMan-MicroRNA-Assays-and-Arrays.html

http://www.qiagen.com/Products/Catalog/Assay-Technologies/miRNA/miScript-Primer-Assays

http://www.exiqon.com/miRNA-qpcr-designer

Hi Richard,

it depends on which miRNA you are going to study. U will be needing a control miRNA like U6snRNA, Now there are lot of providers who actually provides custom made primers. For instances Qiagen SA bio sciences, Invitrogen, Genecoepia etc. Now Quagen uses sybr green prob and invitrogen uses taqman probe. Both of them provides assay for 96 plate and onwards. Also they provide single tube assay. Go to Qiagen SA biosciences website and then in the bottom of the page Go to the "miRNA research " option. There u will find everything u need. Same for invitrogen also.

You could just amplify the pri-miRNA stem loop. It is capped and poly-A tailed like any mRNA, and I have amplified several using nothing but a pair of sequence specific primers designed using Primer3plus. How you approach this will depend on the biological question that you are asking.

Hi

I have successfully used stem-loop PCR. It's quite easy but you need to have the good controls.

https://www.researchgate.net/publication/230581041_Identification_and_characterization_of_miRNAs_and_their_potential_targets_in_flax

If you need some details, feel free to ask.

Article Identification and characterization of miRNAs and their pote...