Polybrene Infection / Transfection Reagent Protocol 〔海外情報〕
Retroviral Infection
- Recombinant retroviral stocks are prepared by adding 5 mL of growth medium with 5% serum to a near confluent monolayer of transfected retroviral packaging cells in a 100 mm plate. After 24 hours the medium is removed and filtered through a 0.45 µm filter.
- Cells to be infected with this recombinant retroviral stock are plated at 500,000 cells per 100 mm plate in 10 mL of complete medium.
- Remove the growth medium from the cells 24 hours later. Infect cells with 2 mL of the viral supernatant (or a dilution of the virus stock into 2 mL) in the presence of 5 µg to 10 µg of polybrene per mL (final concentration). Incubate cells for 3 to 6 hours at 37°C.
- Add 8 mL of complete medium. Three days after infection, split the cells 1:5 into selection medium.
References
- Toyoshima, K. and Vogt, P.K., Virology 1969; 38:414–26
- Coelen, J.R., Jose, D.G. and May, J.T. Arch. Virol. 1983; 75:307–11
