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Team:Freiburg/Content/Notebook/Labjournal
From 2014.igem.org
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| + | <h1>Virale Vectoren</h1> | ||
| + | <h2>21.05.14</h2> | ||
| + | <h3>Transfection/ Virus production</h3> | ||
| + | <p>For virus production Phoenix cells (producer cell line) were splitted (well separated) on 100mm plates. At 70% cell density cells were transfected via PEI transfection.</p> | ||
| + | <ul> | ||
| + | <li>remove medium and refill with 5 ml new DMEM</li> | ||
| + | <li>600 µl transfection mastermix was prepared (8 µg pMIG IRES EGFP, 24 µl PEI, rest OptiMEM)</li> | ||
| + | <li>mastermix was incubated 15 min and carefully drop on the plates</li> | ||
| + | </ul> | ||
| + | <p>Plates were incubated at 37°C. The supernatant after 24 was removed and refilled with 5 ml new DMEM, the supernatant after 48 h was collected (refilled with 5 ml DMEM) as well as the supernatant after 72 h.</p> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/0/08/Igem_logo.png" alt="iGEM Logo"> | ||
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<div id = "sidenav"> | <div id = "sidenav"> | ||
<h1>Notebook</h1> | <h1>Notebook</h1> | ||
Revision as of 12:58, 22 September 2014
Virale Vectoren
21.05.14
Transfection/ Virus production
For virus production Phoenix cells (producer cell line) were splitted (well separated) on 100mm plates. At 70% cell density cells were transfected via PEI transfection.
- remove medium and refill with 5 ml new DMEM
- 600 µl transfection mastermix was prepared (8 µg pMIG IRES EGFP, 24 µl PEI, rest OptiMEM)
- mastermix was incubated 15 min and carefully drop on the plates
Plates were incubated at 37°C. The supernatant after 24 was removed and refilled with 5 ml new DMEM, the supernatant after 48 h was collected (refilled with 5 ml DMEM) as well as the supernatant after 72 h.
