Internal standards for RT-PCR

Alexander Yephremov efremov at mpiz-koeln.mpg.de
Mon Aug 28 12:37:19 EST 2000


David Humair wrote:

> Hi netters,
> I want to quantify the expression (in fact decrease) of an mRNA in
> senescing plants. Regarding at some commercial kits (generally for
> animal cells) people use: beta-actin, 18S RNA, GAPDH as standards. I
> have also heard ubiquitin. Has anybody heard things about that. Does
> anybody work with the QuantumRNA kit from Amplion (competimer technique)
> with plant material?
> Do you know if the expression of these standards decreases with the
> early senescence of the plant cells?
>
> Best regards
>
> David Humair
>
> --
> Dr. David Humair
> Station fédérale de recherches en production végétale de Changins
> Laboratoire de génie génétique
> Route de Duiller
> CH-1260 Nyon
>
> Tél : +41-22-3634419
> Email : David.Humair at rac.admin.ch
>
> The hardness of the butter is proportional to the softness of the bread.

Hi David,

I successfully used the Ambion kit with Arabidopsis. Please find details in
the The Plant Cell, Vol. 11, 2187?2201, November 1999.
None of standards is ideal but 18S RNA is good enough when you deal with
pipetting errors etc. Be aware, that senescence is likely to add a
noticeble source of variation if your mRNA is very sensitive to decay.

Best regards,
Alexander
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