I'm looking for a method that can detect between 3 to 9 miRNAs in a single reaction. I thought of a Taqman (or similar chemistry) assay but can't find a reference for it.
Can you help?
Multiplexing with taqman probes from ABI is not a good idea in my opinion (mind that i've been using them for nearly 4 years now). first reason is that you have to validate each combination with single assays each time, that's time and money consuming. Second reason is that there are other companies that do not use specific primers for RT (Exiqon (finally!) and Qiagen are 2 examples but there are others.
If you have to do a lot of multiplexing shift to sybr assays. I have no experience with them but i i have every intention to shift to exiqon. still have to test their assays but should be good.
Let us know if u try anything working good
Thanks for your input. Can I ask whether you have seen variation in using A taqman multiplex RT? It's just, using the described method above in a few examples I have not seen variation using 2 ul each primer and reducing the water volume in the reaction to compensate.
I understand that he might as well go the exiqon route, but in terms of megaplexing a taqman RT, I can't help but think the conentrations of primers are in highly in excess in the default protocol and ABI sell custom primer combinations anyway.