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Assay for Phage Containing the Beta-galactosidase Gene - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda7.html

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]

Assay for Phage Containing the Beta-galactosidase Gene Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda7.html

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]

Basic Information on In Vitro Transcription - http://www.ambion.com/techlib/basics/transcription/index.html

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

The ability to synthesize RNA in the lab is critical to many techniques.Radiolabeled and nonisotopically labeled RNA probes, generated in small scale transcription reactions can be used in blot hybridizations and nuclease protection assays. This article includes information on: Requirements For Transcription, RNA Phage Polymerases, Template Options: Plasmids, PCR Products, Oligonuclotides and cDNA, Sense or Antisense, Conventional Or Large Scale Synthesis, Products for In Vitro Transcription. - [Read Basic Information on In Vitro Transcription]

cDNA Library Screening Protocol - ZAPII - http://www.people.virginia.edu/~gwl6s/protocols/cdna.html

Category: RNA Protocols: cDNA Libraries Library

cDNA Library Screening Protocol - ZAPII Detailed Protocol. XLI-Blue MRF' and cDNA phage library screening. Dr. Laurie. University of Virginia Health Science Center - [Read cDNA Library Screening Protocol - ZAPII]

Clone Genes From a Phage Library Protocol - http://www.ucsf.edu/micro/faculty/Johnson/protocols/protocol1.html

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]

Column Method for Lambda Phage DNA Preparation - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda11.html

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Column Method for Mini-prep of lambda phage DNA purification from lysates. Donis Keller Lab - [Read Column Method for Lambda Phage DNA Preparation]

Column Method for Lambda Phage DNA Preparation Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda11.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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

Construction of the Sequencing Library Protocol - http://sequence-www.stanford.edu/protocols/library.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol first describes the vector preparation and, then, describes the insert preparation. Vital to have an excellent vector in order to produce a sequencing library. Protocol employs the male-specific coliphage M13 as the sequencing vector. M13 is a filamentous phage with a single-stranded, circular genome. M13 is widely used as a vector because many versions are available commercially and because M13 has certain advantages. - [Read Construction of the Sequencing Library Protocol]

Dewhurst Lab Protocols in Phage Display - http://128.151.62.33/pdf/DEWphage01.pdf

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Dewhurst Lab Protocols in Phage Display. - [Read Dewhurst Lab Protocols in Phage Display]

DNA Ligation Protocol from Takara Shuzo Co. - http://www.hawaii.edu/microbiology/MO/ligation.htm

Category: Molecular Biology Protocols: DNA Ligation Protocols

Insertion of DNA into Plasmid Vectors, Insertion of DNA into l Phage Vectors, Self-Circularization of Linear DNA (Intramolecular Ligation), Linker (Adaptor) Ligation. Alam's Lab Univ Hawaii. - [Read DNA Ligation Protocol from Takara Shuzo Co.]

Gel Assay to Determine DNA Content of Phage Lysates Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda3.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

Growth and Purification of 25-100 ug Lambda Clone DNA Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda13.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promoters - http://www.nottingham.ac.uk/~mbzspd/methods/In_vitro_RNA_synthes.html

Category: RNA Protocols: In Vitro Transcription T7 SP6 T3 Protocols

RNA, in vitro synthesis using T3 T7 promoters. Dr. Dawson Univ. Nottingham. - [Read In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promoters]

Lambda gt22a Screening Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/L1.html

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol for lambda gt22a screening. Includes: T-TYN Media + Mg+2; T-TYN Plates; T-TYN Top Agarose; Phage Dilution Buffer (PDB); 10X Kinase Buffer (-DTT); Calf Thymus DNA. - [Read Lambda gt22a Screening Protocol]

Lambda Phage DNA Quickprep - http://www.unizh.ch/botinst/Cyto_Website/schneitzLab/Methods/DNAIsolation/phageQuickprep.html

Category: Molecular Biology Protocols: Phage Protocols and Methods

Lambda Phage DNA Quickprep with recipes for PSB, STE, l-dil and MgCa buffers and media. Kay Schneitz - [Read Lambda Phage DNA Quickprep]

Lambda plate lysis Protocol - http://www.rpgroup.caltech.edu/~paul/protocols/phage/lambda/plate/

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Phage Protocols - Lambda plate lysis. Rob Philip Group. - [Read Lambda plate lysis Protocol]

Liquid Phage Lysates Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda2.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

Long Term Lambda Phage Storage Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda6.html

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

No special treatment is required to prepare a lysate for the active collection. The following procedure should be used for long-term storage of lambda clones in the archival collections. The phage are diluted in media containing 7% DMSO and frozen at -80 degrees C. - [Read Long Term Lambda Phage Storage Protocol]

Natural History Museum Protocol for EST sequencing from lambda zap phage plaques - http://www.nhm.ac.uk/hosted_sites/schisto/protocols/nhm_est_method.html

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]

Paul's Phage Protocols - http://www.rpgroup.caltech.edu/~paul/protocols/phage/

Category: Molecular Biology Protocols: Phage Protocols and Methods

Paul's Phage Protocols. Many phage related protocols - [Read Paul's Phage Protocols]

PCR based Lambda Î»DNA library screening method - http://www.methods.info/Methods/Library_screening/library_pcr.pdf

Category: RNA Protocols: cDNA Libraries Library

"useful screen phage library non-radioactive, and not need too many clones. Up to 5 clones can be obtained usually within one week." Methods.info - [Read PCR based Lambda Î»DNA library screening method]

Ph.D.â„¢-12 Phage Display Peptide Library Kit - http://www.neb.com/nebecomm/ManualFiles/manualE8110.pdf

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Ph.D.â„¢-12 Phage Display Peptide Library Kit NEB. Applications for Drug Discovery using Phage Display Libraries of Peptides. - [Read Ph.D.â„¢-12 Phage Display Peptide Library Kit]

Phage Display Libraries Kits - NEB - http://search.neb.com/query.html?qt=phage+display&Image1.x=0&Image1.y=0

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Phage Display Libraries Kits - NEB (New England Biolabs) - [Read Phage Display Libraries Kits - NEB]

Phage Media Preparation - http://www.rpgroup.caltech.edu/~paul/protocols/phage/media/

Category: Molecular Biology Protocols: Phage Protocols and Methods

Preparation of liquid medium for phage growth. Rob Phillips Group: Physical Biology of the Cell - [Read Phage Media Preparation]

Phage Plate Stock Lysates Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda5.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

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Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

Lambda Phage Protocol Large DNA Preparation

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.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

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.

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

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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