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

miRNAs and transfection
The miRNAs were designed and synthesized by GenePharma (Shanghai, China). The following Homo sapiens (has)-miR-224 mimics were synthesized: sense, 5′-CAA GUC ACU AGU GGU UCC GUU-3′; antisense, 5′-CGG AAC CAC UAG UGA CUU GUU-3′; negative control: sense, 5′-UUC UCC GAA CGU GUC ACG UTT-3′; antisense, 5′-ACG UGA CAC GUU CGG AGA ATT-3′; has-miR-224 inhibitor: 5′-AAC GGA ACC ACU AGU GAC UUG-3′; has-miR-224 inhibitor negative control: 5′-CAG UAC UUU UGU GUA GUA CAA-3′. miRNA transfection was performed using FuGENE HD transfection reagent (Indianapolis, IN, USA). In brief, 4×105 cells were plated in 2 ml medium in the wells of a six-well plate. Cells were incubated overnight to achieve >80% confluence at the time of transfection. In each well, 5 μl miRNA was added to 100 μl serum-free medium. Separately, 5 μl FuGENE HD transfection reagent was added to 100 μl serum-free medium and mixed gently. The transfection complex was incubated for 15 min at room temperature, added to cells, and incubated for 6 h after which the serum-containing medium was replaced. Following transfection, the cells were incubated for 48 and 72 h prior to qPCR and western blot analysis, respectively.

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Discussion

Papers

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Manufacturer protocol

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

Download manufacturer protocol

Videos

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