Hi all,

Does anyone have a good quantitative RT-PCR protocol for estimating miRNA expression in cells ? I do not want to go back to Northern blots thats all.

Ambion is a great resource for all things miRNA. Here are some useful links for miRNA expression using quantitative RT-PCR.


Ambion Technote 12(4) for mirVana qRT-PCR miRNA Detection Kit and Primer Sets
Quantify MicroRNA Expression by RT-PCR


Ambion Experimental Workflow
miRNA Expression Analysis


Ambion Technote 13(4)
Getting Started with microRNA Expression Analysis Research using Real-Time PCR


Ambion's miRNA Database Search


Ambion Technical Bulletin #508
miRNA (microRNA) Reference List


Ambion
miRNA Research Guide - pdf document




Here are a couple of good references for miRNA expression using qRT-PCR:

Diana O Perkins, Clark D Jeffries, L Fredrik Jarskog, J
Michael Thomson, KeithWoods, Martin A Newman, Joel S Parker,
Jianping Jin and Scott M Hammond. microRNA expression in the prefrontal cortex of individuals with schizophrenia and schizoaffective disorder. Genome Biology 2007, 8:R27


Shingara J, Keiger K, Shelton J, Laosinchai-Wolf W, Powers P, Conrad R, Brown D, Labourier E. An optimized isolation and labeling platform for accurate microRNA expression profiling. RNA. 2005 Sep;11(9):1461-70. Epub 2005 Jul 25.

Thanks a lot. Its been really very helpful joining up with you guys. Wish I had been here sooner.

I'm glad we could help! If you remember, please post some details on the method that worked best for you. It would be very helpful for the rest of the board.

Yup. Soon as my miRNA microarrays go through I'll follow up with PCR and keep updating the thread.
:)

I finally used the Agilent platform for doing the microarray analysis. Validation by qRT-PCR was done using ABI miRNA kits. The results seem to match so far :)

I have been using Taqman microRNA assays from ABI too. Like it a lot!!

Hi yuut.

I was just curious to know what sort of application are you using microRNA assays for?

Hi Tony Rook,

I used microRNA assays to quantifiy the expression levels of several microRNAs during certain biological process.