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Title: PCR gel results: do they reflect gene expression levels of different genes? Post by: mido1770 on Sep 14, 2012 For example, if I ran a PCR on several different genes for the same DNA sample, and then put the products through an agarose gel...will one band for Gene 1 being brighter than Gene 2's band reflect higher gene expression for Gene 1? Or is it impossible to compare. This is semi-quantitative RT-PCR.
Title: PCR gel results: do they reflect gene expression levels of different genes? Post by: midnightxxmal on Sep 14, 2012 Not sure about RT_PCR so here a good website, scroll down to pictures of gels
http://www.ambion.com/techlib/basics/rtpcr/index.html Title: PCR gel results: do they reflect gene expression levels of different genes? Post by: rjedlicka on Sep 14, 2012 Semi quantitative agorose gel results give only a basic idea of expression. But RT-PCR is done on RNA samples with reverse transcriptase, and not DNA.
Title: PCR gel results: do they reflect gene expression levels of different genes? Post by: julibyrd on Sep 14, 2012 Yeah, you said first you were running it on a DNA sample. That's not quantitative. If you're using cDNA, then it could be. At the end, you said it's semi-quantitative. If that's true, then yes. Band intensity should correlate to gene expression, if you did the experiment properly. It'd be better to run it on a real-time machine with fluorescent dyes, if you can. Much more accurate.
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