pCMV-HA2/DKK1

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For transfection of the recombinant vector pCMV-HA2/DKKl, Free-Style 293-F cellswere incubated at 37°C and CO2 volume fraction of 8%. The flasks were shaken at 130 rpm. Before transfection, cell density was adjusted to 1 x 106/mL. Next, 100 pg recombinant plasmid pCMV-HA2/DKK1 and 200 mL Free-Style 293-F cells for liposome transfection reagent were diluted to 3.33 mL with Opti-MEM, allowed to stand for 5 min, and the plasmid and transfection reagents were mixed slowly and reacted for 20 min at room temperature.

Protocol tips

For transfection of the recombinant vector pCMV-HA2/DKKl, Free-Style 293-F cellswere incubated at 37°C and CO2 volume fraction of 8%. The flasks were shaken at 130 rpm. Before transfection, cell density was adjusted to 1 x 106/mL. Next, 100 pg recombinant plasmid pCMV-HA2/DKK1 and 200 mL Free-Style 293-F cells for liposome transfection reagent were diluted to 3.33 mL with Opti-MEM, allowed to stand for 5 min, and the plasmid and transfection reagents were mixed slowly and reacted for 20 min at room temperature.

Publication protocol

For transfection of the recombinant vector pCMV-HA2/DKKl, Free-Style 293-F cellswere incubated at 37°C and CO2 volume fraction of 8%. The flasks were shaken at 130 rpm. Before transfection, cell density was adjusted to 1 x 106/mL. Next, 100 pg recombinant plasmid pCMV-HA2/DKK1 and 200 mL Free-Style 293-F cells for liposome transfection reagent were diluted to 3.33 mL with Opti-MEM, allowed to stand for 5 min, and the plasmid and transfection reagents were mixed slowly and reacted for 20 min at room temperature. Next, 93.3 mL Free-Style 293-F cells (density of 1 x 106/mL) was added to the DNAfectin reagent mixture to a volume of 100 mL. The cells were cultured at 130 rpm at 37°C and with 8% CO2.

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