FuGENE® HD Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Always bring FuGENE HD to room temperature and mix well prior to use.

Upstream tips
Always bring FuGENE HD to room temperature and mix well prior to use.

Publication protocol

Lentivirus production and transduction
Lentiviruses were produced and packaged in HEK293T cells in 6-well format. HEK293T cells
were maintained at ~70% confluency before transfection. FuGENE HD transfection reagents
(Promega) were mixed with 0.5 µg of lentiviral vector, 1 µg of pCMV-dR8.2-dvpr vector, and
0.5 µg of pCMV-VSV-G vector in 100 µl of OptiMEM medium (Life Technologies), and were
incubated for 15 minutes at room temperature before adding to cell culture. Culture medium was
replaced the next day. Supernatant containing newly produced viruses were collected at 48-hour
and 96-hour post-transfection, and filtered through a 0.45 µm polyethersulfone membrane (Pall).
Filtered viral supernatant was used to infect cells in the presence of 8 µg/ml polybrene (Sigma)
overnight for transduction with individual vector constructs. For pooled lentiviral library

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Manufacturer protocol

Download the product protocol from Promega for FuGENE® HD Transfection Reagent below.

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