Team:Freiburg/Protocols/Transfection

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Transfection of Cells

Transfection of Cells

material: chemicals, used kits, …
duration: 30 min

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• The transfection is carried out at a density of 50-70%. The following information refers to the transfection of cells on 10 cm plate.
• Just before the addition of the transfection mix 25 µM chloroquine were added to the medium.
• The amount of DNA used was 12μg. The final volume was 1.6 ml.
• First, the DNA was treated with the appropriate volume ddH2O, which was calculated from the discrepancy of the final volume
• then mixed with other ingredients and 99.2 ul of sterile 2 M CaCl2.
• Then 800 ul 2x HBS were added dropwise while vortexing to the approach.
• The transfection mixture was dropped onto the cells within 10 minutes.

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• 2x HBS (500ml): 8 g NaCl; 0,37 g KCl; 106,5 mg Na2HPO4; 1 g Dextrose; 5 g Hepes (pH 7,05)

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