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Measurement of Cell Adhesion Under Static Conditions - http://www.glycotech.com/protocols/Proto8.html

Category: Cell Biology and Cell Culture: Cell Adhesion Protocols

Many proteins and molecules promote cell adhesion including several cell surface carbohydrate binding proteins. Cell adhesion measurements on 96-well microtiter plate format are difficult due to the shear forces generated by washing the wells. The protocol here introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity. This eliminates uncontrolled shear forces and passage of adherent cells through a liquid/air interface. John L. Magnani~GlycoTech Corporation. - [Read Measurement of Cell Adhesion Under Static Conditions]

MICROARRAY cDNA CLONE GROWTH AND TEMPLATE - http://pga.tigr.org/sop/M001_1a.pdf

Category: DNA Microarray Protocols

This protocol describes clone handling, plate replication, and DNA template preparation in a 96 well format. Hasseman. TIGR Microarray Protocols - [Read MICROARRAY cDNA CLONE GROWTH AND TEMPLATE]

Molecular and Genetic Toxicology Studies on Microbial Mutagenesis - http://www.sri.com/biosciences/bdd/toxic/protocol.html#4

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies on microbial mutagenesis. Includes: G101 Salmonella/Microsome Plate Incorporation Assay (Ames Test), G101-R, G102 Salmonella/Microsome Preincubation Assay (Ames Test), G102-R, G103 Salmonella/E. Coli Reverse Mutation Assay (using four Salmonella and one E. coli strains), G104 Salmonella/E. Coli Reverse Mutation Assay (using four Salmonella and two E. coli strains), G105, G106. - [Read Molecular and Genetic Toxicology Studies on Microbial Mutagenesis]

Mouse Embryonic Fibroblast Culture and Propagation Part II PDF - http://www.wicell.org/uploads/media/Protocols_Part_2_05.pdf

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

WiCell Reserarch Institute. Count MEF Cells, Trypsinize, Split, Freeze, Thaw, Irradiate, Reconstitute, Plate, Feeder. - [Read Mouse Embryonic Fibroblast Culture and Propagation Part II PDF]

MTT Assay for Detection of Cellular Proliferation - http://www.ebioscience.com/ebioscience/appls/AC145.htm#mouse

Category: Immunology: T Cell Protocols

Mouse Protocol: Stimulation of mouse peripheral T cells with plate-bound 145-2C11 monoclonal antibody; MTT assay for detection of cellular proliferation. - [Read MTT Assay for Detection of Cellular Proliferation]

Nematode Dye Filling Protocol - http://www.k-state.edu/hermanlab/protocols/nematode_dye_filling.htm

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for nematode dye filling. Includes: Thumb plate; Microfuge tube; 15 ml tube; Viewing: Big Scope and Dissecting Scope. - [Read Nematode Dye Filling Protocol]

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]

Plate Assay for Carboxypeptidase Y in Yeast Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4161

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol for plate assay for carboxypeptidase Y in Yeast cells. - [Read Plate Assay for Carboxypeptidase Y in Yeast Cells Protocol]

PLATE Transformation Protocol - http://www.med.unc.edu/~hdohlman/lazybones.html

Category: Yeast Protocols: Yeast Transformation Protocols

Protocol for PLATE transformation. - [Read PLATE Transformation Protocol]

Preparation of Seeded NGM Plates For Worm Food Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1558

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol describes how to seed plates with E. coli OP50 for C. elegans cultures. Includes: Bacterial Stock Plate Preparation; Bacterial Broth Preparation; Seeding Bacteria Plates for Culturing Worms. - [Read Preparation of Seeded NGM Plates For Worm Food Protocol]

Preparing Stocks of Bacteriophage {lambda} by Plate Lysis and Elution Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3963

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

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]

Preparing the Aggregation Plate Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4420

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes the preparation of aggregation plates, which will then be used for the production of aggregation chimeras. - [Read Preparing the Aggregation Plate Protocol]

Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos - http://www.mshri.on.ca/nagy/Tetraploid/tetra.htm

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Protocol for the production of completely ES cell-derived fetuses by aggregation with tetraploid embryos. Includes: Recovery of 2-cell stage embryos; Production of tetraploid embryos; Preparation of aggregation plate; Removal of Zona Pellucida; ES cells/ tetraploid embryo "SANDWICH" aggregation; Transfer of embryos. - [Read Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos]

Protocol for Luciferase Assay for In Vitro Detection - http://aesop.rutgers.edu/~lamlab/lucassayprotocol.html

Category: Reporter Gene Assays: Luciferase Assay Protocols

Protocol for luciferase assay for in vitro detection. Protocol includes: Before cell lysate preparation; Cell lysate preparation; Protocol for manual luminometers; Protocol for plate reading luminometer; Preparation of protein assay reagent; Protein standards. - [Read Protocol for Luciferase Assay for In Vitro Detection]

Rapid Analysis of Bacteriophage lambda Isolates: Purification of lambda DNA from Plate Lysates - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3984

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol is used to purify small amounts of bacteriophage {lambda} DNA that 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 Plate Lysates]

Removal of Yeast Contamination from Lymphoblast Cultures Protocol - http://humgen.wustl.edu/hdk_lab_manual/hcc/hcc8.html

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when slightly shaken and lymphocytes will not cluster together as much as normal. If cultures are suspect, a drop of culture can be streaked on a YPD media plate to check for growth of yeast colonies, or a 5 ml sample can be taken to Barnes Diagnostic Center for identification of yeast strain. - [Read Removal of Yeast Contamination from Lymphoblast Cultures Protocol]

Screening a cDNA Library for use with HybriZAP zebrafish cDNA libraries - http://www.cbs.umn.edu/~kclark/protocols/library.html

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]

Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3946

Category: Bacterial Protocols

Bacterial colonies growing on agar plates are transferred en masse to nitrocellulose filters. The spatial arrangement of colonies on the plates is preserved on the filters. After transfer, the filters are processed for hybridization to an appropriate radiolabeled probe while the original (master) plate is incubated for a few hours to allow the bacterial colonies to regrow in their original positions. - [Read Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol]

Screening Bacterial Colonies by Hybridization: Large Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3937

Category: Bacterial Protocols

This procedure is used to plate, replicate, and subsequently screen large numbers of bacterial colonies (up to 2 x 104 colonies per 150-mm plate or 104 colonies per 90-mm plate). - [Read Screening Bacterial Colonies by Hybridization: Large Numbers Protocol]

Screening Bacterial Colonies by Hybridization: Small Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3935

Category: Bacterial Protocols

Protocol used to screen a small number of bacterial colonies (<200) that are dispersed over several agar plates and are to be screened by hybridization to the same radiolabeled probe. The colonies are gridded onto a master plate and onto a nitrocellulose or nylon filter laid on the surface of a second agar plate. - [Read Screening Bacterial Colonies by Hybridization: Small Numbers Protocol]

SNP Genotyping System - http://www.promega.com/tbs/tm053/tm053.pdf

Category: Genetics and Genomics: Genotyping Protocols

SNP Genotyping System. Includes: Protocol for Manual READIT® Assay in Multiwell Plates; Preparation of L/L Reagent; Preparation of Heating Block; CIAP/Exo Treatment; Preparation of Interrogation Probe Solutions; Template Denaturation; Sample Interrogation; Protocol for Manual Plate Reading Luminometers; Protocol for Injecting Plate Reading Luminometers; Protocol for Automated READIT® Assay with the Biomek® 2000 Instrument. - [Read SNP Genotyping System]

Streaking Yeast Stocks Protocols - http://humgen.wustl.edu/hdk_lab_manual/yeast/yeast1.html

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

Some yeast strains are unstable (e. g., small YAC-bearing strains) and need to be repurified by streaking on an agar plate and then verifying the genetic content of the isolated colony before proceeding. In cases where the strain is unstable, plan to streak the cells onto the selective medium to retain the desired stock, (however, most strains can be streaked onto the complete medium, YPD). - [Read Streaking Yeast Stocks Protocols]

Target Specific Whole Cell Assay for Antibacterial Drug Discovery Protocol - http://www.natureprotocols.com/2006/06/23/a_targetspecific_whole_cell_as.php

Category: Pharmacology and Toxicology Protocols: Drug Discovery

Protocol describes a target selective S. aureus whole cell assay that combines agar-diffusion and protein over expression techniques. This agar based two-plate differential sensitivity assay was used to help confirm the newly discovered antibiotic platensimycin inhibited bacterial growth by specifically targeting the essential FASII enzyme FabF6. - [Read Target Specific Whole Cell Assay for Antibacterial Drug Discovery Protocol]

Titering of Bacterial Viruses Protocol - http://biology.clc.uc.edu/fankhauser/Labs/Microbiology/Phage/Phage_Titer.htm

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]

Total Internal Reflection Microscopy - http://www.andrew.cmu.edu/user/dcprieve/TIRM.htm

Category: Microscopy Protocols: Optical Microscopy Protocols

TIRM is a optical technique for monitoring the instantaneous separation distance between a microscopic sphere & a flat plate. Changes in distance as small as 1 nm can be detected. Includes information on: Scattering Intensity I is Related to Elevation h ; apparatus. - [Read Total Internal Reflection Microscopy]

Articles (1-9 of 9)

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.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

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.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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