I would like to make cell lines that overexpress using lentivirus.

First, 293T cells are transfected, but there is a problem that the virus is not produced well.

The target plasmid I use is a third-generation lentivirus with a size of 10kb (negative control) and 11.6kb (including insert).

Therefore, I need to use RRE, REV, VSVG together with the target plasmid to generate virus particles.

When you want to proceed like this, we would appreciate it if you could tell us how to transfect the amount or ratio of each plasmid.

It is confusing whether it is only necessary to consider the amount to be entered or to put it in proportions considering the size of each plasmid.

Also, I wonder if adding PEI or CaCl2 makes a lot of difference in improving the efficiency of transfection.

Thank you.