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Team:Freiburg/Notebook/modeling
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Modeling Notebook | Modeling Notebook | ||
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| - | <p>65000 cells per well were seeded. | + | <p>65000 cells per well were seeded. 24 h before transfection.<br> |
| - | The cells had been transfected at 21:30. | + | The cells had been transfected at 21:30. </p> |
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<img src="https://static.igem.org/mediawiki/2013/8/88/Transfektionschema20.8.png" width="600px"> | <img src="https://static.igem.org/mediawiki/2013/8/88/Transfektionschema20.8.png" width="600px"> | ||
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| - | < | + | <p> |
| - | To generate time dependent data, one well was treated every 6h.<br><br> | + | To generate time dependent data, one well was treated every 6h.</p> |
| + | <p> | ||
| + | t<sub>0</sub>= 01:30<br> | ||
| + | t<sub>1</sub>= 07:30<br> | ||
| + | t<sub>2</sub>= 13:30<br> | ||
| + | t<sub>3</sub>= 19:30<br> | ||
| + | t<sub>4</sub>= 01:30<br> | ||
| + | t<sub>5</sub>= 07:50<br> | ||
| + | t<sub>6</sub>= 13:30<br> | ||
| + | </p> | ||
| - | + | <p> | |
| - | + | Every timepoint the supernant was taken for SEAP measurement and the cells were lysated with RIPA buffer for quantitative western blotting.<br> | |
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| - | + | The standard western blot procedure was done with 20 µl of the cell lysates.<br> | |
| - | + | For quantification of the western blot data, a dot blot was done. <br> | |
| - | + | </p> | |
| - | For quantification of the western blot data, a dot blot was done. | + | |
| - | </ | + | |
Revision as of 17:28, 3 October 2013
Modeling Notebook
65000 cells per well were seeded. 24 h before transfection.
The cells had been transfected at 21:30.
To generate time dependent data, one well was treated every 6h.
t0= 01:30
t1= 07:30
t2= 13:30
t3= 19:30
t4= 01:30
t5= 07:50
t6= 13:30
Every timepoint the supernant was taken for SEAP measurement and the cells were lysated with RIPA buffer for quantitative western blotting.
The standard western blot procedure was done with 20 µl of the cell lysates.
For quantification of the western blot data, a dot blot was done.
