Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 1:1 ratio of DNA to diluted Lipofectamine® 2000 Reagent and Incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C. Then analyze transfected cells.

Protocol tips
Add 1:1 ratio of DNA to diluted Lipofectamine® 2000 Reagent and Incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C. Then analyze transfected cells.

Publication protocol

For transfection, 4 × 104 HEK293 cells were seeded on 96-well plates 24 hours before transfection. A reporter construct containing the 3′UTR- of HNF6 downstream of the Renilla luciferase gene27 and the control firefly (pGL3-CMV) luciferase vector, 100 ng and 0.2 ng, respectively, were cotransfected with 25 nM control miRNA or miRIDIAN miR-Mimics (Dharmacon, ThermoFisher Scientific, Tournai, Belgium) and 1 μL Lipofectamine 2000 (Invitrogen). Luciferase activity was measured 24 hours after transfection using the Dual Luciferase reporter assay system (Promega, Leiden, Netherlands) and a Glomax96 luminometer (Promega).

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Manufacturer protocol

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

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