RNA standards for quantitative PCR

[Martin Offterdinger]

On 31 Jan 1997 09:24:58 GMT, "Beisvåg Vidar"
<Vidar.Beisvag at medisin.ntnu.no> wrote:

>Hi, Netters:
>
>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!
>
>Thanks
>
>Vidar
>
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.
Bye Martin