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Lentivirus Production
From 2008.igem.org
(Difference between revisions)
(New page: ==Virus Production Protocol== Aspirate 3 large dishes of HEK-293 cells. Add 12.5 mL of normal cell media to each dish. Return the 3 dishes to the incubator. In a 15mL tube, add DMEM,...) |
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| - | In a 15mL tube, add DMEM, p148, p149, DNA, and lastly Superfect | + | In a 15mL tube, add DMEM, p148, p149, DNA, and lastly Superfect. Pipet up and down GENTLY a couple of times. Incubate a room temperature for 20 minutes to allow liposomes to form. |
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Carefully stack the plates and return them to the 37C incubator. Wait 40-48 hours, and then proceed to virus harvesting. | Carefully stack the plates and return them to the 37C incubator. Wait 40-48 hours, and then proceed to virus harvesting. | ||
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| + | *p148 and p149 are helper plasmids | ||
Revision as of 01:52, 29 October 2008
Virus Production Protocol
Aspirate 3 large dishes of HEK-293 cells.
Add 12.5 mL of normal cell media to each dish.
Return the 3 dishes to the incubator.
In a 15mL tube, add DMEM, p148, p149, DNA, and lastly Superfect. Pipet up and down GENTLY a couple of times. Incubate a room temperature for 20 minutes to allow liposomes to form.
Label the 3 dishes. After the above mixture has been sitting out for 20 minutes, line up the 3 dishes next to each other and add 1mL of the above mixture dropwise to each plate. Distribute the rest of the solution.
Carefully stack the plates and return them to the 37C incubator. Wait 40-48 hours, and then proceed to virus harvesting.
- p148 and p149 are helper plasmids
