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Team:Freiburg/Content/Notebook/Labjournal
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Revision as of 13:05, 22 September 2014
Virale Vectoren
21.05.14
Transfection/ Virus production
For virus production Phoenix cells (producer cell line) were splitted (well separated) on 100mm plates. At 70% cell density cells were transfected via PEI transfection.
- remove medium and refill with 5 ml new DMEM
- 600 µl transfection mastermix was prepared (8 µg pMIG IRES EGFP, 24 µl PEI, rest OptiMEM)
- mastermix was incubated 15 min and carefully drop on the plates
Plates were incubated at 37°C. The supernatant after 24 was removed and refilled with 5 ml new DMEM, the supernatant after 48 h was collected (refilled with 5 ml DMEM) as well as the supernatant after 72 h.
