发信人: sichou (nini), 信区: Biology
标 题: Re: One question about transfection
发信站: BBS 未名空间站 (Sun Jan 15 00:54:16 2006), 转信

1.it seems to me that it's more likely that the expressed protein is toxic t
o the cells since Lipo2000 transfection is a routine operation in our lab an
d does not cause serious toxicity to 293T,Hela and alot other cell lines.

2.for macrophage, you can't use lipo2000, no way~ Macrophages are extremely
difficult to be transfected. Your boss is right, use AMAXA nucleofection and
the efficiency is quite high >50%.


【 在 bewithu (x-fire in the sea) 的大作中提到: 】
: 细胞是primary human fibroblasts
: Plasmids are pEGFP-X.
: Reagent: lipofectamine 2000
: Method: Add the plasmids & lipo (prepared in opti-MEM) to wells with serum -

: free medium （or full medium). After 4 h of incubation, remove the
: medium, and add new one.
: Results: 1. Mock-transfected cells (only lipo added, no DNA) are not
affected
: at all
: 2. For DNA-transfected cells, more than 60% cells expres GFP. But
: viability is not good. Many floating cells (like dying) have
: ...................


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