For RV-transduction, I used calcium phosphate method.
(+ Chloroquinine during transfection/ polybrene during transduction)
Current my problem is that about 2 days after transduction, most of cells started to die.
I have no idea. Previously, it worked well, but suddenly it didn't work at all.
What I changed were HBSS and DMEM (invitro to lonza), 293T cells and I purified new plasmid DNA (endo free)
And other stocks, CaCl2, Chloroquinine, Polybrene are about 2 year old. (But keep them in -20(cacl2/chloro) and -80(poly))
Since non-transduced cells had better cell viability, so I think transduction may affect the cell viability.
But I have no idea what factors may affect the cell condition and viability in this method.
If you have any idea about this problem, please reply!!