I have calculated the copies/ml of my lentiviral particles using qRT-PCR. Now, to calculate MOI, MOI = plaque forming units of virus for infection/number of cells.
In this case, can I equate PFU/ml to copies/ml and calculate MOI similarly?
the qPCR reads (for example) 100 copies/ul, your eluate will contain 300 x 100 (30 000, 3E4) copies, and this is the same as the number of copies in your pellet, so the viral titer would be 3E4/5E6 per cell or 3E4/1500 per ul
Using qPCR to assess viral copy number will give you a correlation with PFU, but these are not the same measurements. A PFU measurement is a functional assay, and only intact infective virions capable of inducing plaque formation will be counted. In a qPCR you will also detect non-infective virions and free RNA leading you to overestimate the amount of virus.
Hadi Jabbar Alagealy understood, thanks!
Jose Risomar Sousa sure
John Hardy Lockhart yes okay, I see!
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