PEI transfection

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PEI transfection Powered By Docstoc
					                                                                 AG 21.11.2005


PEI
Aldrich   Cat.No. 40,872-7      Lot 03307LC-454       100 ml



Prepare new PEI solution for transfection:

Working solution: 1x (= 0,4 mg/ml PEI)

      Dilute 1ml 20x Stock PEI solution with 19 ml NaCl 300mM
      Aliquot it in 1ml
      Store at 4 °C

For 293T cells:

Prepare DNA solution for 6 wells

   1.: dilute 8 µl = µg plasmid DNA in 150 µl DMEM w/o
   2.: add 24 µl well vortexed 1 x PEI solution
                      vortex immediately and spin down gently
   10 min @ RT
   add dropwise 182 µl to the cells in each well
   don´t change the medium
incubate cells over night at 37 °C and 5% CO2



Prepare DNA solution for p100 dishes

   1.: dilute 24 µl = µg plasmid DNA in 900 µl DMEM w/o
   2.: add 72 µl well vortexed 1 x PEI solution
                     vortex immediately and spin down gently
   10 min @ RT
   add dropwise to the cells in each well
   don´t change medium
incubate cells over night at 37 °C and 5% CO2
Prepare new PEI solution for transfection:
                                                                AG 21.11.2005


for HeLa and C2C12 cells:

PEI pH 10



Prepare DNA solution for 6 wells

   1.: dilute 8 µl = µg plasmid DNA in 150 µl DMEM w/o
   2.: add 32 µl well vortexed 1 x PEI solution
                      vortex immediately and spin down gently
   10 min @ RT
   add dropwise 190 µl to the cells in each well
   don´t change the medium
incubate cells over night at 37 °C and 5% CO2




Prepare DNA solution for p100 dishes

   1.: dilute 24 µl = µg plasmid DNA in 600 µl DMEM w/o
   2.: add 96 µl well vortexed 1 x PEI solution
                     vortex immediately and spin down gently
   10 min @ RT
   add 720 µl dropwise to the cells in each plate
   don´t change medium
incubate cells over night at 37 °C and 5% CO2

				
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