I confused my transgenic mice - some questions about transgenic mice (Oct/30/2007 )
hello everyone:
I have a big question about my transgenic mice. Since previous members has made a transgenic mice which can cause neurodegenerative disorders. And they have did some behavior test and IHC stain, the result was really good- there was some apoptosis in mice brain.
But something wrong with our genotyping. We try for 3 months in genotyping.... but it still cannot get a good PCR band on my gel. We can see mice F0 and F1 generation had some bahavior unnormal (clasping, stun), but we cannot validate they are transgenic mice by molecule biology methods. Is there any good method approaching this difficulty? Thanks a lot!
Jiun Yu
Can you describe how these mice were made transgenic? By stable transfection of some vector? Where do your primers for the PCR bind, what`s the template? Maybe your PCR needs optimisation. If you f. ex. knocked out a gene you could show that it is not expressed anymore, but that really depends on your previous experiments.
I have a big question about my transgenic mice. Since previous members has made a transgenic mice which can cause neurodegenerative disorders. And they have did some behavior test and IHC stain, the result was really good- there was some apoptosis in mice brain.
But something wrong with our genotyping. We try for 3 months in genotyping.... but it still cannot get a good PCR band on my gel. We can see mice F0 and F1 generation had some bahavior unnormal (clasping, stun), but we cannot validate they are transgenic mice by molecule biology methods. Is there any good method approaching this difficulty? Thanks a lot!
Jiun Yu