qPCR for microRNAs - minimal primer length? (Jul/09/2007 )

To test the expression of miRs, many qPCR systems are available, e.g. from applied biosystems, ambion and exiqon. Unfortunately, I want to look at a miR for which no qPCR system is available. So I tried RNAse protection, but that turned out to be not sensitive enough.

So I want to design my own primers for the cDNA synthesis and the qPCR for this miR. Does anybody know the minimum primer lengt for the microRNA-qPRC reaction, either taqman or SyberGreen?

Thanks in advance.

-Theo22-

QUOTE (Theo22 @ Jul 9 2007, 02:11 AM)

Hi

The latest sequence at send to AB was short (50 ). If you use the program done by AB (assays by design file builder) I dont think that you will have some problem

Good luck

DoD

-DoD-

See following linke
http://www.protocol-online.org/forums/inde...99&hl=polyA

-rye-

QUOTE (rye @ Jul 10 2007, 12:22 AM)

See following linke
http://www.protocol-online.org/forums/inde...99&hl=polyA

Thanks!

-Theo22-

I have a mture microRNA sequence and would like to design real time PCR primers, could someone please advice me ho wto procede with this. Please note that that particular microRNA is not yet in database and is novel.
Thanks.

-honeyhappy-

QUOTE (honeyhappy @ Sep 12 2008, 01:32 AM)

Hi, I try quite a lot of known and novel microRNA detection by using Quigen-based qRT-PCR techniqen. This is quite sensitive and good noice signal. Try to visit Quigen website for more information

-Raymondlung-

Thanks It looks promising but using sybr green

-honeyhappy-