Mouse Embryonic Stem Cells Transfection

Mouse Embryonic Stem Cells cultured on a matrigel-coated 100 mm dish have been transfected with GeneCellin HTC. We have used 2x10⁶ cells in a 100 mm dish, 15 µg of plasmid DNA and 30 µL of GeneCellin HTC reagent. We have obtained about 50% of transfected embryonic stem cells 48 h post transfection.

Protocol
1) 2x10⁶ cells were seed on a matrigel-coated 100 mm plate (BD Biosciences) the day before transfection
2) 15 µg of plasmid DNA were diluted into 1 mL of serum free DMEM-F12 medium
3) 30 µL of GeneCellin HTC reagent (BioCellChallenge) were added and the mixture incubated 15 min at room temperature
4) mixture was added onto cells
5) plasmid expression level was monitored 48 h post transfection

Thanks to BioCellChallenge technical support team for assistance.