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