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2-D Polyacrylamide Gel Electrophoresis Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/2DPage.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

96-Well Confirmation Yeast PCR Protocol - http://sequence-www.stanford.edu/group/yeast_deletion_project/96_well_confirmation.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for 96-well confirmation Yeast PCR. Includes: Clonal purification; Generate a master plate (96-well format); Making a frozen backup stock; Confirmation PCR for one Row; ORF Specific Confirmation PCR --> "A-B" primers (upstream junction); Transfer template DNA to multiwell PCR plate; Prepare and dispense master mix for A-B PCR. - [Read 96-Well Confirmation Yeast PCR Protocol]

Antibiotic and Selection Reagent Usage for Resistant E. coli Cells PDF - http://www.atcc.org/pdf/AntibioticTable.pdf

Category: Microbiology: Antibiotics Concentration in Media LB

Antibiotics, ampicillin, choramphenicol, G418, X-Gal and IPTG also stock concentrations. ATCC. - [Read Antibiotic and Selection Reagent Usage for Resistant E. coli Cells PDF]

Antibiotic solutions - http://molbiol.ru/eng/protocol/01_05.html

Category: Microbiology: Antibiotics Concentration in Media LB

Working concentrations and stock solutions for antibiotics, preparation of stock solutions for Ampicillin, Kanamycin and others. Practical Molecular Biology - [Read Antibiotic solutions]

Antibiotics - http://www.unizh.ch/botinst/Cyto_Website/schneitzLab/Materials/antibiotics.html

Category: Microbiology: Antibiotics Concentration in Media LB

Antibiotic stock solutions preparation, Storage conditions for antibiotics, Working Concentrations for high copy plasmids. Kay Schneitz Univ Zurich. - [Read Antibiotics]

Antibiotics. and Antibiotics Stock Calculator Tools - http://molbiol.edu.ru/eng/protocol/01_05.html

Category: Microbiology: Antibiotics Concentration in Media LB

Working concentrations and stock solutions and which antibiotics are light sensitive. Preparation of stock solutions and calculator tool to calculate concentrations for stock solutions. Practical Molecular Biology. - [Read Antibiotics. and Antibiotics Stock Calculator Tools]

Arabidopsis Tissue Culture Stock Solutions and Media - http://wheat.pw.usda.gov/~lazo/methods/lazo/tissue.html

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Arabidopsis Tissue Culture Stock Solutions and Media. Lazo Lab. MS MEDIUM FOR ARABIDOPSIS. B5 MEDIUM FOR ARABIDOPSIS. 10X Micronutrients. - [Read Arabidopsis Tissue Culture Stock Solutions and Media]

Bacterial Media Solutions and Stocks - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid11.html

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

agar, Ampicillin Stock, Alkaline Lysis Solution, SOB, SOC media, and X-gal stock solutions. Donis Keller. - [Read Bacterial Media Solutions and Stocks]

CaCl2 Competent Cell Stock Protocol - http://www.bio.indiana.edu/~chenlab/potocols/CompetentCellStock.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Simple Protocol for CaCl2 Competent Cells. Chen Lab. Indiana University Bloomington - [Read CaCl2 Competent Cell Stock Protocol]

Cajal Body Isolation Protocol - http://www.lamondlab.com/f7cbprotocol.htm

Category: Cell Organelle Protocols: Nucleus Protocols

Cajal Body Isolation Protocol. Protocol includes: Sonication, Removal Nucleoi, Gradient One, Gradient two, Concentration and final enrichment of cajal bodies. Also includes: Making 2.55M sucrose stock and Analysis of the enriched Cajal body fraction. - [Read Cajal Body Isolation Protocol]

Expression Protocol in M9 Minimal Media via T7 Promoter - http://structbio.vanderbilt.edu/chazin/wisdom/labpro/M9.html

Category: Bacterial Cell Culture Protocols: Bacterial Cell Culture Media Protocols

This protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 minimal media begins with preparing a 5x stock solution of M9 salts. Generally, M9 salts contain a nitrogen source in the form of NH4Cl. Since we want to add a labeled nitrogen source, our 5x salts are prepared minus NH4Cl. Standard 5 X M9 Minimal Media salts minus nitrogen source For 1L 5xM9 salts: - [Read Expression Protocol in M9 Minimal Media via T7 Promoter]

Fission Yeast Media Protocol - http://www-rcf.usc.edu/~forsburg/media.html

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Yeast Cell Culture Growth Media Protocols

Protocol for fission yeast media. Includes: YES (yeast extract with supplements) for rich complete media; YSO, for modified rich media; EMM (Edinburgh minimal medium) for minimal selective media; stock solutions to make up minimal media; MB, a very stringent minimal for transformation; Sporulation media; ME (malt extract); SPAS; EMMGlut. - [Read Fission Yeast Media Protocol]

How to Avoid Pitfalls and Keep Out of Trouble in the Neurospora Lab - http://www.fgsc.net/neurosporaprotocols/How%20to%20avoid%20pitfalls%20and%20keep%20out%20of%20trouble.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to avoid pitfalls and keep out of trouble in the Neurospora lab. Includes: Culture, incubation, storage, and disposal; Stock preservation; Spore and colony counts; Making crosses and recovering progeny; Use of heterokaryons for allelism tests; Use of unordered asci to estimate the relation of translocation breakpoints to centromeres. - [Read How to Avoid Pitfalls and Keep Out of Trouble in the Neurospora Lab]

How to Deposit Materials in the Fungal Genetics Stock Center (FGSC) - http://www.fgsc.net/neurosporaprotocols/How%20to%20deposit%20materials%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to deposit materials in the Fungal Genetics Stock Center (FGSC). - [Read How to Deposit Materials in the Fungal Genetics Stock Center (FGSC)]

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]

Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol - http://cyto.mednet.ucla.edu/Protocols/simultta.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Protocol for measuring apoptosis and necrosis by dual laser flow cytometry. Includes: Preparation of stock solutions of HO342, PI, and 7-AAD; Staining for detection of apoptosis; - [Read Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol]

Media and Culture of Yeast Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E5192D47CE9B0776DB7D136C34ED148A&objectid=6673ABA6C2DB2E4E7564EDFA26589256

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

The yeast, Saccharyomyces cerevisiae, has become an important organism in molecular, biochemical, and genetic analysis. The organism has specific requirements for growth under a variety of conditions. The media, both liquid and solid, simple, define, and complex are describe in this unit. Also included are methods for handling, storing, and shipping stock of yeast. - [Read Media and Culture of Yeast Protocol]

Method to Restart Stuck Fermentation - http://www.vinquiry.com/pdf/METHODTORESTARTSTUCKFERMENTATION2003.pdf

Category: Bacterial Cell Culture Protocols: Fermentation Equipment

Method to restart stuck fermentation. Includes: Select and rehydrate active dry wine yeast; activate the rehydrated yeast with nutrients and sugar; start the fermentation and add the stock wine in batches. - [Read Method to Restart Stuck Fermentation]

Neurospora Methods Protocols - http://www.fgsc.net/methods/stanford.html

Category: Fungi Protocols: Neurospora Protocols

Protocols for Neurospora methods. Includes: Standard strains; Crosses; Minimal medium; Color coding of media; Agar substrate for manipulation and isolation; Stock solutions of supplements; Mating-type tests; Preservation of stocks by silica gel; Cleaning of glassware; Control of mites. - [Read Neurospora Methods Protocols]

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]

Plating Bacteriophage Lambda Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3961

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]

Preparation and Loading of Protein Samples for Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4656

Category: Microinjection Protocols: Microinjection of Proteins Protocols

his protocol provides methods for the preparation of protein samples and for loading them into pulled microinjection pipettes. Stock solutions of proteins are thawed, diluted (if desired), centrifuged at high speed to remove aggregates, and kept on ice until loading. Loading into micropipettes can be done using either a "front-loading" or a "backfilling" procedure. - [Read Preparation and Loading of Protein Samples for Microinjection 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]

Preparation of Stock Solutions for FISH Protocol - http://users.ugent.be/~fspelema/neubla/protocol/stock.htm

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for the preparation of stock solutions for FISH. - [Read Preparation of Stock Solutions for FISH Protocol]

Producing Single-stranded DNA with Phagemid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4019

Category: Viral Manipulation Protocols: Phage Protocols

Protocol describes methods to superinfect bacteria carrying a recombinant phagemid with a high-titer stock of an appropriate helper virus and to assay the yield of filamentous virus particles that carry single-stranded copies of the phagemid DNA. The key to success in using phagemids is to prepare a stock of helper virus whose titer is accurately known. - [Read Producing Single-stranded DNA with Phagemid Vectors Protocol]

Articles (1-7 of 7)

Preparing Stock Plating of Bacteria Protocol

A protocol for the preparation of stock plating of bacteria.

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.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

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.

Crystal Violet Stain

Crystal Violet stain preparation method and protocol.

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.

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