Protocol Online logo
Top : Forum Archives: : Real-Time PCR

change in expression of House keeping gene - change in expression of House keeping gene (Aug/12/2006 )

i am getting 10 fold increse in my gene of intrest but house keeping gene also showing 2 fold increse , in real time PCR. can i say that there is 5 fold increse in gene (after normalization of housekeeping gene)

-Rohit Saluja-

Some housekeeping genes such as beta-actin can be altered by certain treatment. So if you are sure that the change in housekeeping gene expression is not caused by treatment you can normalize your GOI to housekeeping gene.


It can depend how harsh or rigorous you examiners/supervisors/referees are. A two-fold change is really not that much. However, when you look at publications you sometimes see people quoting effects due to treatment that are only 1.5-fold changes (to the gene of interest). I think this can be clutching at straws ut it depends on the field. I work in drug discovery and we often see 1000-10^4-fold changes in RNA levels. To us 2-fold changes in house keeping genes are irrelevant.

You might try an alternative housekeeper (if you can afford it) to compare effects. GAPDH, 18 S RNA, TBP...