RNA standards for quantitative PCR
a8803349 at unet.univie.ac.at
Fri Jan 31 06:04:10 EST 1997
On 31 Jan 1997 09:24:58 GMT, "Beisvåg Vidar"
<Vidar.Beisvag at medisin.ntnu.no> wrote:
>I am trying to set up a quantitative PCR assay. I want to use an internal
>RNA standard. This could be made by in vitro transkription of a cloned PCR
>product. My problem is that i want a poly A tail on the 5`of the finaly
>transkript (and I dont know how I could make such a product). Then I could
>add the RNA standard in the mRNA isolating step. I want the standard to
>compeet with the mRNA about the olido dT beeds, who I am using for
>isolation of mRNA. If I can do this, it hopefully will be a good assay for
>quantification of expressed genes with help of PCR.
>I hope some one could help me, or know some one who could give me a hint!
I do not know how to prepare a T tagged standard but I would like to
mention that I have always used total RNA for qt.RTPCR with good
sucess and that I do not htink that preparation of mRNA would be any
advantage, because PCR is that sensitive that you should get a signal
even without preparing mRNA.
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