Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Huh7 cells were seeded into 24-well plates at a density of 5 ×104 cells/cm2 in 1 mL of growth medium (DMEM containing 10% fetal bovine serum, supplemented with 2 mM L-glutamine, 1% nonessential amino acid solution, 100 U/mL penicillin, and 100 μg/mL streptomycin). The cells were grown in a humidified atmosphere (5% CO2, 95% air, 37°C) for 24 hours. Prior to transfection, the medium was removed and the cells were rinsed with phosphate-buffered saline (pH 7.4). The cells were incubated with 0.5 mL of the PCL/DNA complexes at various weight ratios containing 1 μg of pDNA for 24 hours at 37°C in a 5% CO2 atmosphere. Nontreated cells and cells transfected with naked plasmid, PEI/DNA complexes, and Lipofectamine 2000/DNA complexes were used as controls. After transfection, the cells were washed twice with phosphate-buffered saline and grown in culture medium for 24 hours to allow for green fluorescent protein expression. All transfection experiments were performed in triplicate.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine® 2000 Transfection Reagent below.

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