There is some controversy on which to use: L32, GAPDH, etc. Some people say not to use GAPDH or Beta Actin as controls for Q-PCR.
Thanks Gowri. I have heard that too. Just some people whom do QPCR tell me not to used GAPDH or Beta Actin because the RNA expression levels vary in the cell lines.
Hi, I think you should test some genes and then decide which gene(s) could be used as a control! Cheers, Nadine
Thanks for your comment Nadine. I have done this already and there are some fluctuations between the genes. I decided after this to see what the general community thought to see if I could get a consensus.
Ok, perfect world solution, get up to 6 primer sets and decide. Either way, to get a good publication, recent reports are including 2 reference genes, so investing in that early step may prove helpful. Moreover, it really depends on your variables. I was studying the effect on cell metabolism and GAPDH was not recommended. Even though it showed no variability in my cell lines, I compared to 18S and TBP(TATA binding protein). It's context dependent really. Hope this helps!
Jonathon,
The Jo Vandesompele, et. al 2002 paper is still crucial to this discussion:
http://www.ncbi.nlm.nih.gov/pubmed/12184808
- Badges
- Science topic