Detecting miRNA expression ?

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Fizz
Fizz's picture
Detecting miRNA expression ?

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.

Tony Rook
Tony Rook's picture
Ambion is a great resource

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.

Fizz
Fizz's picture
Thanks a lot. Its been really

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

Tony Rook
Tony Rook's picture
I'm glad we could help! If

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.

Fizz
Fizz's picture
Yup. Soon as my miRNA

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

Fizz
Fizz's picture
I finally used the Agilent

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 :)

yuut
yuut's picture
I have been using Taqman

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

Fizz wrote:

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 :)
Tony Rook
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Hi yuut.

Hi yuut.

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

yuut
yuut's picture
Hi Tony Rook,

Hi Tony Rook,

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

Tony Rook wrote:

Hi yuut.

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

John Cage
John Cage's picture
Detectors

Are there some detectors available for this measurement? I saw some on them at https://ecotestgroup.com/ Have you ever tried them before? What can you recommend me?