In article <Stephen_Lasky-0212950741390001 at cis-ts1-slip1.cis.brown.edu>,
Stephen_Lasky at brown.edu (Stephen R. Lasky) wrote:
> In article <49l20s$bq at mark.ucdavis.edu>, ez008413 at bullwinkle.ucdavis.edu> (Gabriel Romero) wrote:
>> > I'm about to start gene expression studies at the transcription level but
> > I have not decided between RT-PCR and Northern analysis as the method to
> > use. I only need to have a yes or no answer for expression initially and
> > then have some idea on the level of expression. How do the methods
> > compare in terms of ease/difficulty in running the experiments, and in
> > terms of amount and quality of information?
> > Thanks for any advice.
> > Gabby Romero
>> RT-PCR is much more sensitive, faster, requires less RNA, and is easy to
> do. It is, however, more difficult to quantitate. You also need to be
> able to design good primers (which is usually not too much of a problem).
>i agree. i'm currently running northerns for a number (about 120) of
candidate genes discovered via RT-PCR because the quantitation is much
easier to do with a northern for this number of clones. so if all you
need is a yes or no answer, RT-PCR will likely be quicker and easier to do
for the aforementioned reasons. however, if you end up having a bunch of
them to quantitate, the amount of troubleshooting involved in quantitative
RT-PCR might be more hassle than it's worth.
Eric C. Anderson
Cell Biology and Genetics
Memorial Sloan-Kettering Cancer Center
1275 York Ave., Box 470
New York, NY 10028
e-anderson at ski.mskcc.org