I am trying to concentrate lenti virus particle supernatant obtained from HEK-293T cells using manually prepared 10% sucrose solution. I tried several times, but did not get any pellet. I measured the viral titre in residue remaining after centrifugation and aspiration of the solution by real time PCR, though it does not have distinct pellet. In this case, the viral titre value is very low.
Hi Sailesh,
If you can't detect your virus anywehre in your tube then you probably have a problem in virus propagation. Do you observe chnges in cell shape (cytopathic effect) or medium color?
Try to follow the attached protocol from Nature Protocols 1, - 241 - 245 (2006)
It seems to be straight forward.
Good luck,
Noam
Hi :I am not sure if do you detect the lenti virus titers.Maybe you need more higher titers virus since you want to purify it.HCVcc is different with lenti virus.So I have no idea the method is ok for your lentivirus.
How many plates are you using.Maybe You can try with more plates(15 cm).