I want to silence a specific gene using siRNA and I never do such experiment before, so I have some basic questions.
1) about transfection and gene silencing control.
After I finish tranfection procedure, I have normal 1ry AB against expressed protein so can I check the gene silencing using this kind of AB.
2) After transfection and make sure that my gene is knocked down, I need to do experiment by 1st trypsinonized the cells, remove it and recultrue it again on my specific experiment conditions (this is my procedure). so each time I do experiments I need to make the transfection experiment first?? I cant have stock of silenced cells, or reculture it in plates and take cells every time I need.
3) do u suggest any comprehensible review or article or book for more information regarding siRNA practical steps