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Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3976

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Removal of the 5'-phosphate groups from the internal termini of bacteriophage {lambda} arms is used to prevent self-ligation and suppress the background of nonrecombinant bacteriophages during cloning. - [Read Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol]

Analysis of Recombinant Bacteriophage M13 Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3996

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A rapid method to analyze the size of the single-stranded DNA of M13 recombinants. - [Read Analysis of Recombinant Bacteriophage M13 Clones Protocol]

Assaying the DNA Content of Bacteriophage lambda Stocks and Lysates by Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3967

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes the DNA content of bacteriophage lambda stocks can be easily and rapidly estimated. - [Read Assaying the DNA Content of Bacteriophage lambda Stocks and Lysates by Gel Electrophoresis Protocol]

Bacteriophage Lamda Plant Genomic Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GenomicDNAPhageLibraryConstruction.pdf

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol for bacteriophage lamda plant genomic library construction. Includes: Preparation of the insert DNA; Pilot Sau3A Digestion; Full-scale partial digestions. - [Read Bacteriophage Lamda Plant Genomic Library Construction Protocol]

Cloning into Bacteriophage M13 Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4018

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Protocol describes three standard methods to construct bacteriophage M13 recombinants: (1) ligating insert DNA to a linearized vector, prepared by cleavage of M13 RF with a single restriction enzyme; (2) using alkaline phosphatase to suppress self-ligation of the linearized vector, and (3) using M13 RF cleaved with two restriction enzymes for directional cloning. - [Read Cloning into Bacteriophage M13 Vectors Protocol]

Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3801

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol describes DNA-sequencing of single-stranded DNAtemplates in reactions catalyzed by Sequenase. - [Read Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol]

Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4086

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage {lambda} pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4086

Category: E Coli Protocols: E coli Protein Protocols

Protocol for expression of cloned genes in E. coli using the bacteriophage lambda pL promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage lambda pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Expression of Cloned Genes in E. coli Using the Bacteriophage T7 Promoter Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3597

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression of cloned genes in E. coli using the bacteriophage T7 promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage T7 promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage T7 Promoter Protocol]

Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Formamide Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3973

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Formamide can be used instead of proteinase K to dissociate bacteriophage {lambda} coat proteins from the viral {lambda}. The procedure is rapid and works best with large-scale preparations of bacteriophage {lambda}. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Formamide Protocol]

Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Proteinase K and SDS Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3972

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

DNA is best isolated from bacteriophage lambda particles by digesting the viral coat proteins with a powerful protease such as proteinase K, followed by extraction with phenol:chloroform. This procedure is easily adapted for use with smaller-scale preparations of bacteriophage lambda. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Proteinase K and SDS Protocol]

Generation of a Library of Randomly Overlapping DNA Inserts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4081

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

Shotgun sequencing of a large segment of DNA involves random fragmentation of the target region into smaller segments that are subsequently cloned into a bacteriophage M13 vector. The goal is to create a library of overlapping clones that provide at least fivefold coverage over the entire length of the target fragment. - [Read Generation of a Library of Randomly Overlapping DNA Inserts Protocol]

Generation of Sets of Nested Deletion Mutants with Exonuclease III Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4078

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The double-stranded DNA of recombinant plasmid, phagemid, or bacteriophage M13 replicative form DNA is digested with two restriction enzymes whose sites of cleavage both lie between one end of the target DNA and the binding site for universal primer. The enzyme that cleaves nearer the target sequence must generate either a blunt end or a recessed 3' terminus; the other enzyme must generate a four-nucleotide protruding 3' terminus. - [Read Generation of Sets of Nested Deletion Mutants with Exonuclease III Protocol]

Growing Bacteriophage M13 in Liquid Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3994

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

Hybridization of Bacteriophage DNA on Filters Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3983

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]

Identifying DNA-binding Proteins in Bacteriophage ambda Expression Libraries Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3785

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes how to identify cloned cDNAs encoding proteins that bind to specific DNA sequences. The methods used are very similar to those used for immunological screening of expression libraries except that the nitrocellulose filters carrying immobilized proteins are screened with 32P-labeled double-stranded DNA rather than with antibodies. - [Read Identifying DNA-binding Proteins in Bacteriophage ambda Expression Libraries Protocol]

Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes - http://omrf.ouhsc.edu/~frank/GT11.html

Category: RNA Protocols: cDNA Libraries Library

Adsroption of Anti-E. coli Antibodies, cDNA Library Screening, Frank Lab. Oklahoma Medical Research Foundation - [Read Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes]

Isolation of High-molecular-weight DNA from Mammalian Cells Using Proteinase K and Phenol Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4036

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Isolation from Mammalian Cells Protocols

Method of choice when large amounts of mammalian DNA are required, for example, for Southern blotting (Rapid Isolation of Mammalian DNA, Rapid Isolation of Yeast DNA, Southern Blotting: Capillary Transfer of DNA to Membranes) or for construction of genomic libraries in bacteriophage {lambda} vectors. Approximately 200 µg of mammalian DNA, 100-150 kb in length, is obtained from 5 x 107 cultured aneuploid mammalian cells (e.g., HeLa cells). - [Read Isolation of High-molecular-weight DNA from Mammalian Cells Using Proteinase K and Phenol Protocol]

Large-scale Preparation of Single-stranded and Double-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3284

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Protocol used chiefly to generate large stocks of double-stranded DNA of strains of M13 that are routinely used as cloning vectors. Large amounts of single-stranded DNA of an individual recombinant may occasionally be needed for specific purposes, e.g., to generate many preparations of a particular radiolabeled probe or to construct large numbers of site-directed mutants. - [Read Large-scale Preparation of Single-stranded and Double-stranded Bacteriophage M13 DNA Protocol]

Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3980

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Pilot ligations and packaging reactions are used to establish the amounts of fragmented genomic DNA and bacteriophage {lambda} arms that yield the maximum number of recombinants. Additional ligation and packaging reactions may then be set up to yield a comprehensive library of genomic DNA. - [Read Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol]

Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries - http://molecularstation.com/protocol-links/admin/dir_links_edit.php?action=N

Category: DNA Protocols: cDNA Library Protocols

The final stage of cDNA library construction involves optimizing ligation of the size-fractionated cDNA to bacteriophage {lambda} arms, followed by packaging and plating of the recombinant bacteriophages. - [Read Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries]

Markers for Pulsed-field Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3222

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol fopr markers of pulsed-field gel electrophoresis. Markers for pulsed-field gel electrophorsis can be generated by ligation of linear monomers of bacteriophage {lambda} DNA (48.5 kb) into a nested series of concatemers. This procedure yields a series of concatemers that contain up to 20 tandemly arranged copies of bacteriophage DNA. - [Read Markers for Pulsed-field Gel Electrophoresis Protocol]

Ph.D. Peptide Display Cloning System - NEB - http://www.neb.com/nebecomm/TechBulletinFiles/techbulletinE8101.pdf

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Ph.D. Peptide Display Cloning System - NEB. The Ph.D.â„¢ Peptide Display Cloning System facilitates the display of custom peptide libraries on the surface of bacteriophage M13 as coat protein fusions, creating a physical linkage between each displayed pep - [Read Ph.D. Peptide Display Cloning System - NEB]

Picking Bacteriophage lambda Plaques Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3962

Category: Viral Manipulation Protocols: Phage Protocols

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

Picking Single Phage Plaques Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda1.html

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]

Articles (1-2 of 2)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

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