X-tremeGENE™ HP DNA Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Do not aliquot X-tremeGENE HP DNA Transfection Reagent; store in the original glass vials.

Upstream tips
Do not aliquot X-tremeGENE HP DNA Transfection Reagent; store in the original glass vials.

Publication protocol

HepG2 cells were transfected with the miR-425 promoter-Luc construct or an empty pGL3-Basic vector without the miR-425 promoter sequence (Empty-Luc construct) using X-treme GENE HP DNA transfection reagent (Roche,7 Indianapolis, IN) according to the manufacturer’s protocol. A construct directing constitutive expression of renilla luciferase was also transfected to control for transfection efficiency. Twenty-four hours after transfection of the constructs, the cells were cultured overnight in medium containing 1mM glucose (Sigma) and then treated with 30mM glucose for 6h. After 6h of the glucose treatment, cells were harvested, and firefly and renilla luciferase activities in cell extracts were measured using the Dual Luciferase Reporter Assay System (Promega, Madison, WI) according to the manufacturer's protocol.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for X-tremeGENE™ HP DNA Transfection Reagent below.

Download manufacturer protocol

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