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Detection of Ubiquitylated Proteins in Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4615

Category: Yeast Protocols: Yeast Protein Protocols

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting. - [Read Detection of Ubiquitylated Proteins in Yeast Protocol]

Determining Plating Efficiency in Yeast: Direct Method - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4185

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Direct method for determining efficiency in yeast. The plating efficiency of a strain is a measure of the percentage of cells in a culture that are capable of forming colonies (colony forming . - [Read Determining Plating Efficiency in Yeast: Direct Method]

Determining Plating Efficiency in Yeast: Indirect Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4184

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Protocol for the indirect method of determining plating efficiency in yeast. - [Read Determining Plating Efficiency in Yeast: Indirect Method Protocol]

Direct PCR from Whole Yeast Cells Protocol - http://www.duke.edu/web/ceramide/protocols/0003.html

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

Protocol for direct PCR from whole Yeast cells. - [Read Direct PCR from Whole Yeast Cells Protocol]

Directed Yeast Two Hybrid Screening for MEKK1 Interaction Partners Protocol - http://www.natureprotocols.com/2007/01/22/directed_yeast_two_hybrid_scre.php

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol utilizes the Y190 yeast strain, that characteristically turns from white to red during growth due to ademutation. Baits were subcloned into the pAS1cyh2 vector, and Preys subcloned into pACT2. - [Read Directed Yeast Two Hybrid Screening for MEKK1 Interaction Partners Protocol]

DNA Preparation from 10-mL Yeast Culture Protocol - http://www.duke.edu/web/ceramide/protocols/0002.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol for DNA preparation from 10-mL Yeast culture. - [Read DNA Preparation from 10-mL Yeast Culture Protocol]

DNA-Binding Protein Purification with Dynabeads. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2671937

Category: DNA Protocols: EMSA DNA-Protein Protocols

Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.]

Drosophila Cornmeal, Dextrose and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/dextrosefood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, dextrose and yeast medium. - [Read Drosophila Cornmeal, Dextrose and Yeast Medium Protocol]

Drosophila Cornmeal, Molasses and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/molassesfood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, molasses and yeast medium. - [Read Drosophila Cornmeal, Molasses and Yeast Medium Protocol]

Drosophila Cornmeal, Sucrose and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/harvardfood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, sucrose and yeast medium. - [Read Drosophila Cornmeal, Sucrose and Yeast Medium Protocol]

Drosophila Cornmeal, Sucrose, Dextrose, Yeast and 2-Acid Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/caltechfood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, sucrose, dextrose, yeast and 2-acid medium. - [Read Drosophila Cornmeal, Sucrose, Dextrose, Yeast and 2-Acid Medium Protocol]

Drosophila Sucrose and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/neurofood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila sucrose and yeast medium. - [Read Drosophila Sucrose and Yeast Medium Protocol]

EMS Mutagenesis of Yeast Protocol - http://www.fhcrc.org/science/labs/hahn/methods/genetic_meth/ems_mutagenesis.html

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol for EMS mutagenesis of yeast. - [Read EMS Mutagenesis of Yeast Protocol]

Ethyl Methane Sulfonate (EMS) Mutagenesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4180

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol describes mutagenesis of yeast with ethyl methane sulfonate (EMS). It causes approximately 40-70% cell death in most haploid laboratory strains, a level of cell killing that is commonly used in mutant hunts with haploid strains. - [Read Ethyl Methane Sulfonate (EMS) Mutagenesis Protocol]

EXAMINING THE FUNCTION OF PROTEINS AND PROTEIN NETWORKS WITH THE YEAST TWO-HYBRID SYSTEM - http://proteome.wayne.edu/Update.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Interactor Hunt Protocol. Testing whether a bait activates transcription. Screening a library for interactors. Finley Lab. 1997. - [Read EXAMINING THE FUNCTION OF PROTEINS AND PROTEIN NETWORKS WITH THE YEAST TWO-HYBRID SYSTEM]

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]

FM 4-64 Labeling of Yeast Vacuole Membranes Protocol - http://www.colorado.edu/MCDB/odorizzilab/FM464labeling.pdf

Category: Microscopy Protocols

FM 4-64 is a lipophilic styryl dye and a vital stain: it fluoresces only in living cells, so cells cannot be fixed then stained nor stained then fixed. You must stain and observe living cells. FM 4-64 does not permeate cell membranes but, instead, intercalates into the plasma membrane is then taken into the cells by endocytosis. - [Read FM 4-64 Labeling of Yeast Vacuole Membranes Protocol]

Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting - http://www.axis-shield.com/densityhome/optiprep/S29.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Fractionation of (a) vacuolar and subvacuolar vesicles and (b) vacuole and cytoplasm-to-vacuole targeting (Cvt) vesicles from yeast spheroplasts in a pre-formed discontinuous iodixanol gradients. Protocol includes: Formation of yeast spheroplasts; Isolation and vesiculation of the vacuoles; Separation of the vacuolar and subvacuolar vesicles; Separation of vacuoles and Cvt vesicles from a yeast spheroplast lysate. - [Read Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting]

Fractionation of Yeast Membranes in Pre-Formed Continuous Iodixanol Gradients - http://www.axis-shield.com/densityhome/optiprep/S32.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Membrane Isolation Protocols

This protocol is concerned with the use of iodixanol gradients in an analytical mode to study the membrane localization of a particular protein or function. Continuous gradients are best suited to this task. One of the protocols described in this protocol starts with a discontinuous gradient, but since the gradient is centrifuged at 174,000g for 16 h it will become continuous by diffusion. - [Read Fractionation of Yeast Membranes in Pre-Formed Continuous Iodixanol Gradients]

Frozen Yeast Transformation Protocol - http://www.umanitoba.ca/faculties/medicine/biochem/gietz/Frozen.html

Category: Yeast Protocols

This Protocol allows You to prepare Frozen Yeast cells that are competent for transformation after thawing. - [Read Frozen Yeast Transformation Protocol]

Gateway-Compatible Yeast One-Hybrid Screens Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/28/pdb.prot4590

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Protocol for gateway-compatible yeast one-hybrid screens. - [Read Gateway-Compatible Yeast One-Hybrid Screens Protocol]

Gene Disruption via PCR Protocol - http://www.fhcrc.org/science/labs/gottschling/yeast/pcrko.html

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

Protocol for gene disruption via PCR. - [Read Gene Disruption via PCR Protocol]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4562

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This procedure capitalizes on the asexual/sexual reproductive cycle of yeast cells. Laboratory yeast strains can be maintained indefinitely by asexual reproduction as one of two haploid mating types, MATa or MAT{alpha}. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4563

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes how to handle arrayed yeast using robotics. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array:]

GFP Fixation in Yeast Protocol - http://www.ciwemb.edu/labs/koshland/Protocols/MICROSCOPY/gfpfix.html

Category: Yeast Protocols: Yeast Staining Protocols

Protocol for GFP fixation in Yeast. - [Read GFP Fixation in Yeast Protocol]

Articles (1-5 of 5)

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.

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.

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.

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