miRNA real time PCR - miRNA real time PCR (Oct/31/2007 )
it is so expensive to buy the real time PCR kit for miRNA analysis, it includes
mirVana™ qRT-PCR miRNA Detection Kit，SuperTaq™ Polymerase、mirVana™ qRT-PCR Primer Set for Normalizatio、mirVana™ qRT-PCR Primer Sets。
I can only afford the miRNA and U6 primers, can anybody tell me whether there is a way to replace other reagent with cheap reagent or ordinary probes?
I want to see whether my overexpression of miRNA worked or not and try to find a miR-21 low or no expressed cell line.
Personally, I would not get the Ambion system. We tried it and could never get consistent results and spent a lot of time trying to optimize it. We got the Applied Biosystems TaqMan probes and they worked perfectly first time. I think they're more expensive, but it will work. When you buy the RT kit for the miRNA qPCR from ABI, you can get extra reverse transcriptase separately, since this is the reagent that I use up first.
How about the public probe libary (PPL) from Roche?
It seems inexpenssive and effective Anyone used this ,please make a comment on it ,OK?