I am performing minimum MOI experiment. The volume I calculated at moi 0.01 is 0.0115. Since the selected volume is very small, I made a 1:100 dilution and put 1.15ul of phage lysate into 98.5 buffer.
But when I similarly dilute moi 10 (the volume to be added is 11.5ul), it will get 1150ul of phage lysate, which I think is incorrect.
How to dilute the two, because I have to choose 100ul from both of them.
You can do serial dilution a pic the right phage quantity at the same volume. For example if you hqve a stcok of 2E8 pfu you orepqra rikute -1; -2;-3;-4;-5;-6;-7. If you need 2E3 phage for ml than you pick the volume 2E6 dilution one mililitet. In other hande in E5 you will have 2E3 phage for ml in 100ul volume.
You just subtract from 100 the amount of your phage dilution you will add and then use that value for the buffer.
So if you add 1.15ul of phage you will add 98.85 ul of buffer
If you add 11.5ul of phage then you use 89.5ul of buffer
However please note that pipettes are really not accurate at those tiny volumes. So if you want accurate results I would not attempt to pipet volumes less than 5ul for any part of the experiment.
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