I am performing real time to quantify relative gene expression in treated versus untreated samples. How do I plot the ct values in form of gene expression as compared to expression of a housekeeping gene?
the biorad guide is excellent in my opinion
http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_5279.pdf
Once you got ct values there are many methods to study relative gene expressions or fold change like delta delta Ct or delta ct etc find here the detailed information embedded link for relative quantification. http://gene-quantification.com/pfaffl-rel-quan-book-ch3.pdf
Hi Mansi
What real-time machine are you using? With quite a few of them you're able to account for this in the experimental run and it will perform relative expression for you.
Regards,
Nathaniel
Hello Nathaneil, I am using stepone plus real time machine from applied biosystems
If you want the formulas, please consult Hellemans et al., Genome Biology, 2007. If you want software to do the calculations for you, please check Biogazelle's qbase+ software.
https://www.biogazelle.com/qbaseplus
Article qBase relative quantification framework and software for man...
hello Mansi
Stepone Plus software can do these calculations for you. it gives the fold change in form of RQ. however it is good to go through the basics and understand how it is calculated. the link provided by Raghavendra is good. you should go through it.
Late comer here!
If you do R, please try my MCMC.qpcr package. It does all calculations AND plots things very nicely. Grab the step by step tutorial from my RG page...
Thank you all for your contributions. I find them very useful in my PhD research.
