I performed a biopaning and elute the phage using 0.1M Gly-HCL pH2.2, after nuetralizaiton , phage titer was done. And I plate the TG1 ,infected with eluted library, on Kana and Amp plate , incubate at 37degree overnight.
at Day2, there is 300 colonies on Amp plate , but more than 1000 colonies on Kana plate.
I am confused now about that what is that growed on Kana plate, M13KO7 or anything else?
If you are using M13K07 as a helper phage then that is most likely what you are getting on the kanamycin plates.
Michael J. Benedik Thanks Michael, I was doing biopaning , and after elution, titration of eluted library was performed, infected TG1 were plated on Kan+ and Amp+ plate, respectively. Theoretically, it should be only Amp+ plate that colony grow well. Is there any possible that M13KO7 plasmid and phagemid were package into a single phage particles?
Not likely that both were packaged in the same particles. You could test that if you wish, pick 10 colonies from the amp plates and see if they are Kan resistant, and do the reverse as well. Mostly likely there is just so much M13K07 around that you got plenty of those along for the ride. On the other hand, don't worry about it. Just work with your Amp resistant colonies and see if you can find the ones you want.
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