semi-quantitative RT-PCR

"Vladimir Grubor" <forwards nospam at hgmp.mrc.ac.uk
Thu Nov 28 09:17:22 EST 2002


Hi there,

I have some experience with Semi-quantitative RTPCR. I am not sure about the
kit, since I don't use any..You need to realise that the fact that you use
PCR means that any difference in amplification efficiency between different
amplicons will be increased by several orders of magnitude by end of 30-40
cycles. Using semi-quantitative RTPCR, you can only compare single amplicon
from different RNA preps from different tissues/life stages/etc, with use of
household gene which *should* have same level of expression as RNA
loading/PCR conditions control.

Best of luck,

Vladimir


"Stefi~" <xxxgifalone at tin.it> wrote in message
news:eP8F9.211$fa.16608 at news1.tin.it...
> Hi everyone,
> I need to relative quantify the amount of a mRNA during embryonic and
> metamorphic development in the common toad.
> I've recently bought a kit from Ambion: 18S Internal Standard, in which
the
> 18S rRNA is co-amplified together with my mRNA of interest.
> Is there anyone of you who has already used this kit? Any particular
> suggestion? Tips and tricks?
> Thanx so much in advance.
>
> Stefano
> xxxgifalone at tin.it (remove xxx)
>
>





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