Amplify XCX15 random peptide library for epitope mapping - (Sep/18/2007 )
I am trying to amplify an old stock of XCX15 mer random peptide phage library using TG1 cells. I initially tried an old stock of K91 cells: the cells should be at an OD550nm of 2.0 for infection with this phage stock. The problem is the cells grow well until OD 550nm reach 1.6 after 7hrs! and thenI had to leave them two night at 37deg shaking:it was then that the OD reached 2.2! This is not good! I am now trying TG1 cells, still I encounter the same problem. Inputs will be aprreciated.
Why you not centrifuge the cells and resuspend them in a less volume?
I have constructed several cDNA libraries by lambda phage vector, I always culture the host cells to an OD600 nm at between 0.5-1.0, then centrifuge them and resuspend them with 10 mM MgSO4 at half volume of the initial volume.