After I transfer 293 cells with lipofectamine 2000 and Pac I digested Vector, almost all of 293 cells float.
The follow is my protocol:
1. plate cell in 6 well plate with DMEM without antibiotics. After 24h,cells reached 90% confluent.
2. prepare DNA-LF2000 mixture strictly according to manufacture`s manual. 4uL DNA and 10uL pofectamine2000 was added per well.
3. remove cells medium with 2mL serum free DMEM and add DNA-LF 2000 complexes gently.
4. After incubate for 6h, chang medium with fresh complete DMEM.But after 12h all of cells had float.
how can I decrease the LF2000 toxic effect and modify the protocol?
thanks a lot:)
Edited by hhhhyyyy, 03 August 2004 - 06:46 AM.