My PI wants me to do a phage library amplification as the first step to do phage display. How to do the phage library amplification, and what is the concept behind? Please let me know your thought, thanks!
Phage library amplification is essentially to increase the volume of the phage library you have. Let us say you have 1ml of the library, but you can amplify it so that you now have 10ml or 100 ml so that you have enough of the library to work with.
The problem with amplifying a library is that if some clones in the library don't grow quite as well as others (for whatever reason) then these will become more underrepresented or even lost as you continue to amplify the library repeatedly. So that is a concern, but the vast majority of the clones will amplify fine.
One very important thing is that you need to know the initial "complexity" of the library, in other words how many independent clones were there when the library was made. And when you amplify you need to be sure that you always work with numbers larger than the initial complexity.
In other words, if the initial library had a complexity of 106 (independent clones) and the titer of the current library is 109 per ml, then 1 ml of the library would be 1000-fold more than the library complexity. But 1ul would only be equal to the library complexity and hence probably not really include all the various representatives in the library. I usually like to use at least 10-100 fold in excess of the initial complexity to ensure I don't lose things.
So you need to know (1) complexity of the library and (2) titer of the current library stock in order to calculate how much of the existing library you need to use in order to amplify it.
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