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From 2014.igem.org

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.