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Phage display, library dilution

phage display library dilution phage titer

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#1 matey23



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Posted 17 April 2015 - 10:58 AM


I have a question regarding dillution of phage library for 1st (and subsequents) round of panning. I received a kit containing a 100 ul of library with titer 10^13 pfu/ml (diversity 10^9). The manual says: 
Dilute a 100-fold representation of the library (e .g ., 10^11 phage for a library with 10^9 clones) with 1 ml of TBST (100 μl if using microtiter wells) . Pipette onto coated plate and rock gently for 10–60 minutes at room temperature.
I am going to coat an 8 well of microtiter plate with my target. What volume of phage library I should take to 1st round of selection? The whole library (100 ul) diluted to 1 ml with TBST? 
If I took only a 10 ul of the library I would get a 10^11 pfu as a total titer. This is a 100 copy of each variant. Then, if I diluted 10 ul of the library to 800 ul (100 ul/well) I would get approximately a 10 copy of each variant per well. Is it enough for panning?

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