Phage, lysates

Patterson, T. A., and Dean, M. (1987). Preparation of High Titer Lambda Phage Lysates. Nucleic Acids Res 15 6298. 

It is necessary to make high-titer stocks of the recombinant phage for the generation of stocks of phage to work with on a day-to-day basis, and also for the preparation of DNA for sequencing or for the preparation of protein lysates from recombinant phage. A relatively quick and easy method for the preparation of high-titer lysates is described below. 

It can utilize either purified DNA (both super-coiled and linear molecules), broth culture of a plasmid-infected bacteria, or the lysate of a phage-infected bacteria culture as the source of the target sequences. 

NaCl, 10 mM MgSO, 0.01% gelatin), precipitated in the presence of PEG and DNA is isolated by phenol-chloroform extraction as described by Maniatis et al. (7). Alternatively, phage from the plate lysate can be obtained by immune precipitation using a polyclonal antibody directed against lambda phage particles (Lambda-Sorb, Promega Biotech). 

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