I recently bought Lenti-vpak packaging kit - packaging plasmids from Origene and need to use it for transfection of Lentiviral shRNA constructs into X293T cells. Could you suggest the recommended ratio of these packaging plasmids to the shRNA constructs to be used in the Lipofectamine 2000 protocol?
Shanshan Wu Howland
It should be 1 ug shRNA plasmid to 1.2 ug packaging plasmids according to the specs sheet. This is for one well of a 6-well plate. You don't want to use Origene's MegaTran transfection agent provided in the kit? For lipofectamine, you typically want 1:2 to 1:3 ratio. So maybe start with 5.5 ul lipofectamine for the 2.2 ug total plasmid (1:2.5 ratio) for a 6-well plate.
http://www.origene.com/assets/documents/HuSH/Lenti-vpak-app-guide.pdf
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