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Lentivirus Harvesting
From 2008.igem.org
PRINCETON IGEM 2008
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Virus Harvesting Protocol
Set centrifuge to 4C
Filter media with cells from virus production plate using 0.2um or 0.22um filter to sterilize. The virus will go through the filter; any cells / other biological material will not.
When working with live virus, bleach everything – cell culture plates, pipette, filter… and then put them into the orange-bagged bucket.
Sterilely pour filtrate into a polycarbonate ultracentrifuge tube.
Exquisitely balance (fill another tube with water to get them the same weight).
Set centrifuge to 25.50 rotor ID and 50,000g for 2h30m. Load polycarbonate ultracentrifuge tubes and start.
Aspirate bleach from cell culture plates, tubes, etc.
Wipe down hood first with bleach and then with ethanol.
After centrifugation is completed:
Resuspend pellet in 150ul PBS (phosphate-buffered saline).
Incubate for 20min in 4C
Aliquot into 10 tubes (~15ul each).
Flash-freeze in dry ice. Store in -80C
