Housekeeping Genes

Olivier Gandrillon Gandrillon at maccgmc.univ-lyon1.fr
Tue Jul 17 04:10:33 EST 2001


Hi

Except if you are (un)lucky, most of the genes you're gonna test are not 
gonna show up differentially expressed. Then it's just a matter of how 
many different genes are necessary to gain confidence. Two or three 
unrelated (like one house keeping, one transcription factor and say one 
membrane bound receptor: the message here is: ANY gene will be fine...), 
the three of them being exactly overlaping in light cycler using the 
same input of RNA (yes, this IS possible), I would say you're pretty 
confident any other differential will be *true*.

Hope this helps

Olivier

In article <2816803747.994954249 at pc026089.rlh.liv.ac.uk>, 
C.W.Herbert at liverpool.ac.uk (Chris Herbert) wrote:

> Hi there
> 
> I am currently carrying out some Real-Time PCR experiments using a Roche 
> Lightcycler on tumour and non-tumour tissue samples. As this is a very 
> sensitive technique I require reliable housekeeping genes to use as 
> internal standards for these experiments.  It has been suggested that I 
> use 
> GAPDH, but I am reluctant to use this as it has recently been suggested 
> that GAPDH levels increase dramatically in times of cellular stress.  I 
> have used Beta-Actin as an internal control for my experiments so far but 
> wish to validate my results by checking that levels of cDNA are 
> normalised 
> using a second or third housekeeping gene.
> 
> Can anybody suggest other housekeeping genes that I can use?
> 
> Many thanks for your help.
> 
> Chris.
> 
> 
> ---




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