FuGENE® HD Transfection Reagent

DNA transfection Mammalian cells - Immortalized cell lines HEK293

Experiment
DNA transfection Mammalian cells - Immortalized cell lines HEK293
Product
FuGENE® HD Transfection Reagent from Promega
Manufacturer
Promega

Protocol tips

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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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA transfection Mammalian cells - Immortalized cell lines HEK293 using FuGENE® HD Transfection Reagent from Promega.

Paper title
Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM
Full paper
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Manufacturer protocol

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

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