pGEn2-GFP-FcgRIIIa

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add plasmid DNA (300 μl of 0.5 μg/μl) to the culture using micro-pipette and mix by gentle manual shaking. Add PEI (900 μl of 0.5 μg/μl) to the culture, mix by gently shaking. Add 3.8 ml of fresh culture medium, from step 3.1.3, above and transfer flask to the incubator shaker for 24 hr.

Protocol tips
Add plasmid DNA (300 μl of 0.5 μg/μl) to the culture using micro-pipette and mix by gentle manual shaking. Add PEI (900 μl of 0.5 μg/μl) to the culture, mix by gently shaking. Add 3.8 ml of fresh culture medium, from step 3.1.3, above and transfer flask to the incubator shaker for 24 hr.

Publication protocol

"Vigorously resuspend the pelleted cells by pipetting up and down using 10 ml of fresh culture medium and transfer to the flask with fresh transfection culture medium (prepared in step 3.1.3 above). Screw the cap on the vented cell culture flask and move the flask to the incubator (as set in 1.1.1) for 15 min-1 hr with shaking.

During this time, dilute the plasmid DNA and PEI stocks using fresh culture medium (using the 5 ml volume withdrawn from step 3.1.3 above) to a final concentration of 0.5 μg/μl. See sample calculations in Supplementary Materials to determine the amount of DNA and PEI. Transfer the culture flask with the dispersed cells into the biosafety cabinet.

Add plasmid DNA (300 μl of 0.5 μg/μl) to the culture using micro-pipette and mix by gentle manual shaking. Add PEI (900 μl of 0.5 μg/μl) to the culture, mix by gently shaking. Add 3.8 ml of fresh culture medium, from step 3.1.3, above and transfer flask to the incubator shaker for 24 hr."

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Manufacturer protocol

Download the product protocol from Adam Barb, The Roy J. Carver Department of Biochemistry, Biophys for pGEn2-GFP-FcgRIIIa below.

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