Biology Forums - Study Force

Hello I am really lost at the moment, can anyone explain to me the semi-quantitative RT-PCR process?

I don't understand how, by comparing a housekeeping  genes band intestity (after PCR cycles and gel electrophoresis), we can measure the target gene expression?

What will the differences or similarities mean and say about the target gene?

This paper describes it well (attached).

In addition, this article has a good explanation: http://stanxterm.aecom.yu.edu/wiki/index.php?page=Semi_-_quantitative_RTPCR

Thanks so much, the only thing that is bothering me now is...

If I am using specific primers from my reference, and different specific primers for my target (in the same sample) what type of RT-PCR would this be? Competitive or another?

Thanks again

Why don't you just use a regular PCR protocol?

Here are some sources that describe when to use competitive.

1) http://www.uni-ulm.de/~dkaufman/rt-pcr.html
2) http://www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Pierce/realtimepcr.htm
3) http://www.scielo.br/pdf/bjmbr/v36n12/4730.pdf
4) http://www.gene-quantification.com/competitive-rt-pcr.pdf

Thanks for the links,
I was given a protocol; find specific primers for GADPH and specific primers for CD29 beta integrin.
Then theoretically conducted RT-PCR and ran them on a gel.

Because the primers are different, competitive PCR seems unlikely