GAPDH Tissue Distribution - (Sep/19/2009 )
I'm an Honours student completing my studies at Deakin University, Victoria, Australia. Lately, I've been experiecing a lot of trouble-shooting with the tissue distribution of my gene of interest (GOI) and GAPDH. See, after amplification, I use the conventional electrophoresis technique to visualise my GOI, and this is fine. I get good results. However, the moment I add GAPDH to the gel, the bands for my GOI disappear! On a plus side, though, all bands for GAPDH are ubiquitously expressed.
Does anybody know what I could be doing wrong? I'm kind of in the panic zone at the moment considering I only have 4 weeks to produce results and a thesis!
Take a longer exposure picture or expose twice, once with your GOI and once with GAPDH and then overlay images in an image editing program. You will need to have run ladder with your samples for the second to work.
The other option is that GAPDH is the same or similar size to your GOI.