Baculovirus Expression - Sonicate infected cells? (Nov/19/2005 )

I'm infecting Sf9 cells with a bacmid and am about to harvest the initial transfection. I am wondering if I would make sense to sonicate the cells to extract the cellular virus prior to spinning down the cells?
Thanks in advance!

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Greetings !!!
Generally it's not recommended. Recombinant baculoviruses give high titer in media, and thus you don't have to sonicate the cells. Another thing is that sonication would release whole cytoplasm into media. After centrifugation your lysate would cotain many useless compounds. I've never done sonication with Sf-9's and results were OK.

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hi,

i agree w/ hantavirus.... your infected media contains the viruses. once cells (insect-sf9, sf21, etc) are infected, protein will be either secreted or in the cytoplasm. use sterile technique to separate the media from cells via centrifugation and save them both. infected cells can be lysed and purify the protein of interest and as for the media, find out what is the pfu (plaque forming unit) and decide if it needs higher viral titer. it will ususally need 2 to 3 weeks to achieve optimal pfu from the initial transfection.....

good luck,

steven

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