I need to enumerate a large number of viral samples. Typically, I would run each concentration of my sample in all 12 wells of a 96-well plate but in the literature I see 4 or 8 wells per concentration being used. Using fewer wells would save significant time and resources but I don't want to do so at the expense of my data. Can someone point me towards a resource that provides insight into what an increased sample size per concentration does for the strength of my data?
Also, should the TCID50 be run in triplicate for each sample (three dilution series, three plates)? When I think of a plaque assay or bacterial enumeration I would typically do one dilution series then plate in triplicate. Would this be different and why?
All I can tell you is that during my PhD, I used to work with 96 well plates, with 6 replicates + 2 controls (first and last line), doing 12 serial dilutions. If you have an idea of the TCID50, you can still do 2 viruses on the same plate with 6 serial dilutions. I have never done triplicate of my plates. I think 6 replicates in one plate is enough.
I don't really answer your question, but if I could have reduced the amount of plates, it would have avoid me a bad tendonitis!
Good luck!
If your pipettors have been calibrated, and you change tips with each dilution, three replicates will suffice. Many people do not change pipette tips which will cause major variations in titers. There should be no differences in titer between replicates if you are very careful in the pipetting.
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