I am using phagmids as a silencing technique, so there is need of co-transformation of both phagmid and m13 plasmids for core production and insertion of Phagmid into m13 phage core, to avoid co-transformation, can we use another strategy? like isolate both phagmids and core and make the core to pickup phagmids outside the host cell.
In-vitro phage packaging is obsolete and very expensive. The protocol you are using is better, quicker and efficient.
Best,
I am not aware of any in vitro packaging extracts for filamentous phage, but see if you can adapt this to your purposes:
Article Polycos vectors: A system for packaging filamentous phage an...
Hope it helps!
Alejandro Martin You are absolutely right. There is no commercial kit for in-vitro packaging of filamentous phage like M13. That was the reason I mentioned it is an obsolete and very expensive protocol. It's good you shared a very good article. I hope Sayed will definitely get the clue from it.
Packaging of filamentous phages is very complex and involves protein secretion mechanisms across the inner membrane of E. coli. So it is not conducive to in vitro packaging. Have you considered just using a helper phage for infection rather than co-transformation, for example using M13K07.
Sir Michael J. Benedik, I am actually using M13 helper Plasmid (M13KO7), but the problem i am facing is co-transformation even sequential transformation, single transformation has been done for both plasmids(M13KO7 and pAB.001) in DH5alpha, i am assuming that it may be the problem with antibiotic Conc. but i used different Conc for both antibiotics (Amp and Kan), unlucky i didnt get it. any suggestions for co-transformation and antibiotic and plasmids compatibility? thank you all
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