Team:Freiburg/Protocols/Transfection
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Transfection of Cells
Transfection of Cells
material: chemicals, used kits, …
duration: 30 min
- 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)