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RV- transfection-transduction problem


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#1 likepjh



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Posted 10 February 2015 - 04:11 PM



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!!








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