plaques Protocols

Category: Cloning Protocols: Baculovirus Protein Expression Protocols

Protocol for amplification of Baculovirus plaques and preparation of high titer virus stocks. Includes: Primary Amplification; Secondary Amplification. - [Read Amplification of Baculovirus Plaques and Preparation of High Titer Virus Stocks Protocol]

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

This assay is used when working with phage vectors carrying the beta-gal gene. If the cloning event disrupts a normally functional copy of the gene in the vector the resulting plaques would appear clear in the assay. If the phages contain a functional beta-gal gene they will form blue rings around their plaques. Any strain which is not an overproducer of beta-gal will work as indicator host bacteria; a single chromosomal copy of the gene is not a problem. - [Read Assay for Phage Containing the Beta-galactosidase Gene]

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

This assay is used when working with phage vectors carrying the beta-galactosidase gene (often used for immunological screening). If the cloning event disrupts a normally functional copy of the gene in the vector the resulting plaques would appear clear in the assay. If the phages contain a functional beta-galactosidase gene they will form blue rings around their plaques. Any strain which is not an overproducer of beta- galactosidase will work as indicator host bacteria. - [Read Assay for Phage Containing the Beta-galactosidase Gene Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

A silver stain to demonstrate neurofibrillary tangles, nerve fibers and senile plaques in Alzheimer's disease. The nerve fibers are sensitized with a silver solution.  The sections are treated with ammoniacal silver, and then reduced to a visible metallic silver. - [Read Bielschowsky Technique for Senile Plaques Protocol]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Protocol for cloning genes from a phage library. Includes: Titer and plate out phage; Lift plaques onto filters and prepare them for screening; Make a probe; Hybridize the probe to the filters; Wash the filters and expose to film; Purify putative plaques; Excise plasmid from the desired phage. - [Read Clone Genes From a Phage Library Protocol]

Category: Viral Manipulation Protocols: 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]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol for methenamine silver technique for senille plaques. - [Read Methenamine Silver Technique For Senile Plaques Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol for modified thioflavin for senile plaques amyloid. - [Read Modified Thioflavin for Senile Plaques Amyloid Protocol]

Category: RNA Protocols: cDNA Libraries Library

(by PCR of phage insert, cycle sequencing using ABI dye terminator kits and automated sequencing on an ABI 373A) Ian Ridgers - [Read Natural History Museum Protocol for EST sequencing from lambda zap phage plaques]

Category: Molecular Biology Protocols: Phage Protocols and Methods

Plating Lambda Phage to Generate Plaques. Elution of Phage. Phage DNA Extraction. Dr.Wurtzel. Dept. Biological Sciences, Lehman College. - [Read Phage Titer]

Category: Viral Manipulation Protocols: Phage Protocols

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

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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

Category: Viral Manipulation Protocols: Phage Protocols

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]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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

Category: PCR Protocols: PCR Cloning Protocols

In this protocol sequences cloned in standard bacteriophage or plasmid vectors are amplified in PCRs containing primers targeted to flanking vector sequences.  The amplified fragments can be analyzed by gel electrophoresis, DNA sequencing, and/or restriction mapping.  Many colonies or plaques can be assayed simultaneously. - [Read Rapid Characterization of DNAs Cloned in Prokaryotic Vectors Protocol]

Category: RNA Protocols: cDNA Libraries Library

Procedures: Titer the library to determine the concentration of infectious phage, Plate and lift the phage particles, Hybridize the immobilized cDNA with labeled probe, Pick positive plaques. Karl Clark U. Minn. - [Read Screening a cDNA Library for use with HybriZAP zebrafish cDNA libraries]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

An expression library constructed in a bacteriophage {lambda} vector is plated on an appropriate E. coli strain in the absence of isopropylthio-ß-D-galactoside (IPTG). After 2-4 hours, the plates are moved to 37°C (to stabilize any fusion proteins that are temperature sensitive), and filters impregnated with IPTG are laid on top of the developing plaques. - [Read Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol]

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Plaques formed by M13 bacteriophages or bacterial colonies transformed by plasmids carrying specific mutations can be detected by hybridization, using a radiolabeled oligonucleotide that forms a perfect duplex with the mutant sequence. Hybridization is carried out under conditions of low stringency that allow the radiolabeled oligonucleotide to anneal to both mutant and wild-type DNAs. - [Read Screening Recombinant Clones for Site-directed Mutagenesis by Hybridization to Radiolabeled Oligos]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol for senile plaques and neurofibrillary tangles. - [Read Senile Plaques and Neurofibrillary Tangles Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

Category: Viral Manipulation Protocols: Phage Protocols

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

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]