Internal standards for RT-PCR

Frauke Naumann fnaumann at scan.genetik.uni-koeln.de
Thu Aug 17 11:37:10 EST 2000


David Humair schrieb in Nachricht <39940CC2.16A05259 at rac.admin.ch>...
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?


Hi David,
why don't you use an artificial reference that binds to the same primer pair
that you are using in the RT-PCR? Then you wouldn't have to worry about
decreasing expression during senescence. Try Biotechniques 14, 244-249
(1993) - a collegue of mine used this in combination with the RT-PCR-Kit of
Promega (JBC (2000) 275 (9): 6447-6452)
hope that is useful
Frauke
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


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