GADPH does not express - (Oct/24/2013 )
I'm in trouble with GADPH expression. I don't know why GADPH does not express in my samples now, although GADPH always express well and steadly in my previous experiments. I used the same samples ( from ovarian cancer cell lines) which used to express GADPH well but they haven't expressed GADPH now. I tried to used hotstart pcr Premix of bioneer and nTaq hot DNA polymerase of enzynomics, none works. I think my PCR machine has no problem because other genes still express well. I suggested my primer had problem, I ordered new one but GADPH expression can not be detectable again. I have never experienced this problem. Frankly, I don't know what's wrong? Anyone can suggest me what's my problem and how I deal with it?
Thank you so much
Are any other genes being amplified from your ovarian samples? It is possible that your samples have been degraded. Try your primers with freshly extracted/prepared DNA.