Hi,
So, am new to the use of lentivirusses for transduction, so this might be a pretty basic question.
I have made a lentivirus to express my protein of interest and an empty vector that lacks the coding region as a control. I am planning an experiment now to trial different concentrations of my construct to find the maximum efficiency.
Can I trial my main construct and then use the same concentration of the control ?
Or would the appropriate way of doing this be to trial both constructs and then for a final dose select the dose that yields comparable transduction efficiency ( I assume the efficiency might vary between constructs ?)
Any help is greatly appreciated !
Cheers,
Hi Pieter
Whatever MOI you use for your lentivirus needs to be compared to a control (the empty backbone) of the same concentration.
If you do not do this it may skew your results when you test your gene's expression using RT-qPCR.
You technically could do it the other way if you are testing a large variation of MOIs, but if you are only doing 3 or 4 you may as well get the most accurate results possible...empty backbone can still affect cell growth etc.
Good luck
Rachel
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