Hi all, I am currently doing the northern blot using Ambion NorthenMax kit, and also use pTRI-GAPDH mouse for control probe preparation. The weird thing is the GAPDH signal is perfect but my interested genes gave no signal at all...
For my probes targeting interested genes, I used PCR generated DNA template for probe preparation, is that OK? I just directly added T7 promoter at the 5' of my sequence. All the probes are prepared using Roche DIG labeling system.
Hope you guys can give me some suggestions and advice, thank you so much.