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Lentivirus infection of human PBMCs

human transfectio

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#1 immunorange

immunorange

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Posted 04 November 2011 - 10:33 AM

I have viruses for shRNA KD of a gene. I now need to KD the gene in primary human PBMCs, for which I need to infect them with the lentivirus. I know a lot of labs use spin infection.
I stimulate cells with aCD3/CD28, and add polybrene, spin infect after 24hrs. (However this killed all my cells)

1- When do I need to infect them after stimulation to achieve maximum transfection efficiency?
2- Can I not spin them and just add virus+polybrene?
3- Do I need to wash after 3-5hrs of infection? (I dont really want to disturb the blast formation..)

Any help would be appreciated.

Thank you.




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