We are trying to do a negative selection on the library before going to positive selection on our target.
Unfortunately, the phage barely adsorbed into the different cells we used. Anyone have any suggestions?
At what stage are you trying to do the negative selection? I would only try this in the second selection cycle or higher, but not with the naive library. Also, depletion of phage with cells not expressing target receptor would likely not be significant in the first selection round.
Thanks for your answers I will add some more details:
I'm now trying to do the negative selection on attached cells starting from the naive library.
I was using different type of normal primary culture cells. I dilute my library in PBS (10µl in 1 ml). 3 washes in PBS and then incubation for 1h and 30min for 3 times in 3 walls. The incubation is in the incubator then doing the titration and getting no reduction or almost no reduction
Thanks in advance
Bc.R
Thanks for the answers, is there a chance that the peptides are not working from some reason?
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