Experiments NOT working.
I am in my lab now, my experiments are still not working (I am now trying to find out why), and I am pretty fed up now, that's why I am writing this...It is definitely true that, for people working in a lab, their mood is determined by how their mice, cells, bacteria, antibodies and PCR behave.
Nothing seems to be working well recently. Even that nest of baby mice that I tried to keep till P15 died because their mum stopped feeding them after I took 3 baby mice out at P5.
Now I have to wait till I can get hold of a pregnant mouse and wait another 19 days. If I couldn't get those P5 tissue to stain, I will seriously cry.
I don't think it is the antibody's problem; most probably the problem lies with the tissue sample. My antigen seems to be either killed or masked after I kept them for a long time in sucrose to get rid of ice crystal artifacts - and it mysteriously doesn't work on paraffin. The level of expression is not distinctly high, making staining very difficult... I have done it before though, and I have proven its existence using Western blot, but I think pretty soon I have to get a Q-PCR running in order to confirm it at the RNA level. Just to make sure that I am not seeing things like mouse IgG (same size as my protein) - Q-PCR cannot cheat me.
Those bacteria that I transformed are acting funny also.
If anyone has ideas, please please please tell me! Anything would be good; I will know whether it is ingenious or idiotic after I tried it :P
Labels: Life
