psPAX2 and pCMV-VSV-G plus trasnfer RATIOS?

[owl.crow]

Dear All,


Could somebody please help me by telling me the ratios used for the follwoing lentivirus plasmid
:

 psPAX2 and pCMV-VSV-G  plus either LentiCRISPRv2 or LentiGuide-PURO  


I would really really appreciate some comments!

Thank you very much!

[Neville Sanjana]

Hello,

I have commonly seen both 4:3:2 or 4:3:1 ratios. (TransferPlasmid : psPAX2 : pVSVg )  I personally use 4:3:2 and find it works well. For a detailed virus production protocol, please see Chen*, Sanjana* et al. (Cell, 2015) and, in particular, the Pooled Guide-Only Library Cloning and Viral Production section in the Supplementary Methods.

Hope that helps,

Neville

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