Viral Culture Protocols

Protocol for CHAPOL lineage analysis. - [Read CHAPOL Lineage Analyis Protocol]

Mini-prep method for lambda phage DNA purification from lysates. - [Read Column Method for Lambda Phage DNA Preparation Protocol]

Protocol for the electroporation for sindbis virus production. - [Read Electroporation for Sindbis Virus Production Protocol]

A crude lysate gel assay can be performed to roughly quantitate the DNA in lysates. This is often a valuable time saving step to determine if the phage yield is sufficient to warrant continuing the procedure. - [Read Gel Assay to Determine DNA Content of Phage Lysates Protocol]

Protocol for the generation of B95-8 EB-Virus. - [Read Generation of B95-8 EB-Virus Protocol]

Most manipulations with M13, including preparations of viral stocks and isolation of single- and double-stranded DNAs, begin with small-scale liquid cultures that are infected with an M13 plaque, picked from an agar plate. - [Read Growing Bacteriophage M13 in Liquid Culture Protocol]

There are essentially three parts to this protocol: 1. growth of at least 5x10e8 pfu phage to provide an inoculum growth of a larger liquid lysate that will produce about 5x10e12 pfu; 2. concentration and purification of the phage, and; 3. DNA preparation. - [Read Growth and Purification of 25-100 ug Lambda Clone DNA Protocol]

Protocol for lambda methods media and solutions. - [Read Lambda Methods Media and Solutions Protocol]

5 ml liquid lysates are prepared when a small amount of DNA from a large number of lambda clones is needed. The lysates can be made using 10- 20 ul of a stock lysate or a 100-fold amplified phage "macroplaque" as the inoculum. - [Read Liquid Phage Lysates Protocol]

To prepare phage lysates to be used for small or large scale phage DNA preps. This method usually produces lysates with titers of 2-8x10e10 pfu/ml. - [Read Phage Plate Stock Lysates Protocol]

Protocol for the isolation of single bacteriophage lambda plaques to be used for making lysates. - [Read Picking Single Phage Plaques Protocol]

Bacteriophage M13 forms turbid plaques on lawns of male strains of E. coli. - [Read Plating Bacteriophage M13 Protocol]

Bacteriophage {lambda} particles are recovered from bacterial lysates by precipitation with polyethylene glycol at high ionic strength. - [Read Precipitation of Bacteriophage {lambda} Particles from Large-scale Lysates Protocol]

In this protocol, a bacterial lysogen is constructed from a recombinant bacteriophage {lambda} encoding a fusion protein of interest. The resulting lysogenic colonies are induced to synthesize the fusion protein, which is then isolated in preparation for functional and biochemical analyses. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda} Lysogens]

This rapid method is used to screen bacteriophage {lambda}gt11 clones for the production of immunodetectable fusion proteins. After optimizng the conditions of infection and induction, the method can be used to produce preparative amounts of a fusion protein. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda}: Lytic Infection]

A reliable method to prepare plate lysates and to recover infectious bacteriophages by elution from the top agar. - [Read Preparing Stocks of Bacteriophage {lambda} by Plate Lysis and Elution Protocol]

High-titer stocks of bacteriophage {lambda} are easily prepared by infecting small-scale bacterial cultures. - [Read Preparing Stocks of Bacteriophage {lambda} by Small-scale Liquid Culture Protocol]

This protocol is used to purify small amounts of recombinant DNAs cloned in robust strains of bacteriophage {lambda} such as {lambda}gt10, {lambda}gt11, {lambda}ZAP, or ZipLox. The DNAs are suitable for use as substrates for restriction enzymes and templates for DNA and RNA polymerases. - [Read Rapid Analysis of Bacteriophage {lambda} Isolates: Purification of {lambda} DNA from Liquid Cultures]

Phage are streaked onto a medium to obtain an independent isolate prior to preparing a new lysate. This is done to reduce the likelihood of working with lysates which have become contaminated, and/or have accumulated mutations. - [Read Streaking Lambda Phages Protocol]

When an individual bacterial virus grows in a bacterial host suspended in a top agar lawn, its progeny infect and lyse the surrounding host cells. This causes the appearance of a "hole" or plaque in the otherwise homogeneous bacterial lawn. Since each plaque represents a single virus, the number of viruses in the aliquot added to the plate is equal to the number of plaques which appear. - [Read Titering of Bacterial Viruses Protocol]

To transfer phage DNA to nylon membranes so that a phage library can be screened by hybridization. - [Read Transfer of Phage DNA from Plaques to Nylon Membrane or Nitrocellulose Protocol]