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Briefly, female flies 3 5 days old were anesthetized on ice and injected intra-abdominally with ∼100 nl of Dulbecco's modified Eagle medium containing virus at the selected concentration.
For transduction, targeted cells were incubated with medium containing virus for 24 h.
Medium containing virus was removed from cells and replaced by 2% complete medium.
Medium containing virus was collected following 24 and 48 h post transfection and filtered through 0.45-μm filters.
To transduce ASCs, conditioned medium containing virus with dsRed and neomycin resistance was added to ASCs at 70%% confluence.
K562, HL60 and Jurkat cells were incubated with medium containing virus supplemented with 8 μg/ml polybrene for 24 h.
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Filtered medium containing viruses carrying PDGFRβ was used to infect PDGFRα+/+ β−/− MEF.
After washing and addition of the virus medium (not containing virus in case of mock-infection) cells adapt to the new cultivation conditions.
Medium containing recombinant virus was recovered two days after transfection and 10 ml of virus-containing medium was used to transduce 2.5×106 Hcells cells for 24 h.
ϕCRIP or PA317 medium containing infectious virus was harvested (using appropriate containment procedures) and frozen.
The culture medium containing recombinant virus was harvested, filtered and stored at 4°C.
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CEO of Professional Science Editing for Scientists @ prosciediting.com