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

Cell culture and transfection
Cell lines were obtained from ATCC (http://www.atcc.org/). HEK293 cells were grown in Modified Essential Medium with Earles salts (Invitrogen) supplemented with 10% fetal bovine serum, 1× non-essential amino acids, 50 U/mL penicillin, and 50 μg/mL streptomycin (all from Invitrogen). Neuruo2a cells were cultured in Dulbecco's Modified Eagle Medium (Invitrogen) supplemented with 10% fetal bovine serum, 50 U/mL penicillin, and 50 μg/mL streptomycin. For transient expression, cells were plated on a 35-mm glass-bottom dish (Iwaki; Chiba, Japan) and transfected with expression vectors (2 μg) using the FuGENE HD transfection reagent (Promega) and incubated at 37°C for 24–48 h in the medium.

To obtain HEK293 and Neuro2a cells stably expressing human OPN5, transfected cells were selected in the medium containing G418 (1 mg/mL; Invitrogen). G418-resistant cells were then grown in the medium, and OPN5 gene expression was analysed by reverse-transcription polymerase chain reaction (RT-PCR).



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Manufacturer protocol

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

Download manufacturer protocol

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