Phage Protocols

Using hybridization, it is possible to identify a single recombinant that carries the desired target sequence on a filter that carries the imprint of 15,000 or more plaques. - [Read Hybridization of Bacteriophage DNA on Filters Protocol]

General method to pick and store plaques. - [Read Picking Bacteriophage lambda Plaques Protocol]

Protocol describes a method for generating isolated plaques from a stock of bacteriophage lambda. Each plaque derives from infection of a single bacterium by a single bacteriophage particle. Because each plaque contains the progeny of a single virus particle, the bacteriophages derived from a single plaque are essentially genetically identical to one another. - [Read Plating Bacteriophage Lambda Protocol]

Protocol describes methods to superinfect bacteria carrying a recombinant phagemid with a high-titer stock of an appropriate helper virus and to assay the yield of filamentous virus particles that carry single-stranded copies of the phagemid DNA. The key to success in using phagemids is to prepare a stock of helper virus whose titer is accurately known. - [Read Producing Single-stranded DNA with Phagemid Vectors Protocol]

Protocol is used to identify and isolate recombinants containing DNA sequences of interest. - [Read Transfer of Bacteriophage DNA from Plaques to Filters Protocol]

Protocol for transfer of P1 clones by electroporation. - [Read Transferring Bacteriophage P1 Clones between E. coli Hosts Protocol]

Protocol describes methods for recovery and purification of recombinant clones of bacteriophage P1 or PAC DNAs from bacteria. Because of their large size, these DNAs are sensitive to shearing forces and must be handled carefully. This protocol generally yields P1 DNA that works well as a substrate or template in enzymatic reactions. - [Read Working with Bacteriophage P1 and Its Cloning Systems Protocol]

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

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