Hi, i want to produce a high titer of lentivirus using psPAX2, pVSVg, and third plasmid (my GOI cloned) but I am confused about what molar ratio will be best to get higher titer of lentivirus. i read the literature, people used different molar ratios, some used 1:1:1, 8:1:9, and so on. Does anybody know the best molar ratio of the above plasmids?
Thanks
The ratio probably has to do with the size of the third plasmid. The psPAX2 plasmid has the polymerase so the larger the third plasmid the more polymerase will be needed to reach the same titers.
I have the same problem; doing bibliography you can find very different ratio, total DNA, number of cells to transfect, times of supernatant recovery.... my conclusion is that it depends on the vectors (promoter are different) cells (HEKT or HEK or CHO...) maybe on the growth media used (serum? antibiotics? ) and on the product used to transfect (lipo, calcium, PEI.....): the only solution is probably to try different conditions and see what is working in your lab.... (and generally once it works in your hand then you can made variations to improve....)
good luck
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