Gandrillon at maccgmc.univ-lyon1.fr
Tue Jul 17 04:10:33 EST 2001
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
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
> 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
> using a second or third housekeeping gene.
> Can anybody suggest other housekeeping genes that I can use?
> Many thanks for your help.
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