I have estimated the virus titre to be 10 6.5 TCID50/50 µl. I want to know that how much virus I needed to add to infect 25cc flask at 1000 TCID50?
thanks Andrei but my question is I have already calculated the TCID50 of the virus. but now I want to know the volume of virus I have to add in a 25 cc flask (3*10^6 cells) at 100 TCID50. the titre of the virus is 10^6 TCID50/µl
The 10^6 TCID50/05 µl, it means that if you dilute virus stock in 10^6 times you will obtained 1 infection dose of virus. You need 1000 infections dose. Thus you need will prepare the volume which needed for 25 cc flask. (about 2 ml solution contain 1000 infections dose of virus).
Using a TCID50 of 10^6/ul - you have 3x10^6 cells.
Therefore 1 TCID50 = 3ul, 100TCID50=300ul
However, PFU is usually around half of the TCID50, therefore if you want to use plaque forming units, i.e. infectious replicating virus you would have to double this volume. (this is only an estimate assuming you are using the same cells and nothing is added to the assay that will inhibit virus replication).
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