I. Product Description
This lentivirus concentrated reagent is a 5× concentrated solution, which can be used to simply and rapidly concentrate lentivirus particles. This reagent is easy to operate. Just mix the 5× concentrated solution with the lentivirus supernatant in proportion and incubate for a period of time. Then centrifuge at normal speed to obtain concentrated lentivirus particles. The recovery rate is as high as over 90%, and the virus titer can be increased by 100 times.
Ii. Operating Steps
Collect the cell culture supernatant and filter it through a 0.45μm filter membrane to remove cells and debris.
Note: For membrane filtration, cellulose acetate or polyethersulfone (PES) membranes with low protein adsorption should be used. Never use nitrocellulose membrane (NC).
2. Mix the lentivirus supernatant with the concentrated solution in the kit in a volume ratio of 4:1 (4 parts of supernatant and 1 part of concentrated solution), mix well and let it stand at 4℃. Mix once every 30 minutes for a total of 3 times. Incubate at 4℃ for 2 hours or overnight.
Note: Extending the incubation time can increase the recovery rate. Lentivirus particles can be stably stored for several days at 4℃.
Centrifuge at 3.4 ° C and 4000g for 30 minutes.
4. Carefully remove the supernatant. Do not shake the tube vigorously. Generally, white sediment can be seen (sometimes the sediment is not visible).
5. Resuspend the virus particles with DMEM or PBS of 1/10 to 1/100 of the initial volume (original supernatant volume), carefully pipette until uniform, and resuspend the precipitate.
The resuspended virus was aliquoted at 50µl per tube and stored in a -80 ° C refrigerator.
Note: Lentivirus cryopreservation solution should avoid repeated freezing and thawing, otherwise the virus titer will be reduced.