We use 293T cell line for packaging of lentivirus and i need to detect the viral titre. I usually depend of RFP as a control to detect the efficiency of transfection.
who knows a helpful method or methods to detect the exact titre of lentivirus before transduction of my target cell line.
Thanks
Hi Amr
You can run a qPCR or an ELISA for p24.
Take a look at this article. It has the pros and cons of each one.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1534021/
qPCR or p24 ELISA isn't really all that useful for titering a lentiviral vector as these numbers don't tell you the number/concentration of infectious particles in your lentiviral prep.
Does your lentiviral vector also express a fluorescent protein or an antibiotic selection marker? If the former you can use flow cytometry, if the latter you can determine the number of antibiotic-resistant colonies after transduction of your target cell line.
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