Hi, everyone! Recently I'm collecting lentivirus. After package in 293T cells, I take the medium, centrifuge at 1000rpm for 5 min to pellet the cells, then take the supernatant and filter by 0.45um filter. After filtering, I add 1/3 volume of concentration liquid to concentrate the virus at 4℃ overnight. I want to use 1500g, 45min to centrifuge the virus down. Will that speed(1500g) and time (45min) work? If it work, will I see virus particle after centrifugation? If not, what speed and time I need to use to centrifuge?
I could suggest you to use virus-collection protocol which I attach bellow. Conditions of viral harvest are available on the 3rd page. Hope this would be useful for you.
Good luck.
Hi, Kristina, thanks for your suggestion and protocol! I'm trying the method my Lab established now. If it works, then everything is OK, if not, I'll talk to my supervisor and try the protocol you provide. Really thanks for your help!
Hi Siyuan,
We centrifuge over night in a refrigerated bench top tissue culture centrifuge (5000 rpm for 20 hours) - we find that it is kinder to lentivirus vector compared with the ultracentrifuge.
BW, Simon
Hi, Simon,
Thanks for your suggestion. Yesterday I used 1500g, 45min to centrifuge the lentivirus, and it worked! I find that if the concentration liquid (Lenti-X concentrator, ClonTech) is added, then this conditions(1500g, 45min) can work.
Thanks all the same!
With the CloneTech concentrator solution, you can centrifuge low speed, relatively short time. Our standard condition (without addition of concentration solution) is 12000g 4°C overnight.
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