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Preparation of cell lysates from yeast using glass beads vortexing - http://www.embl.de/ExternalInfo/protein_unit/draft_frames/protocol_database/Purification/ec_glass_beads.html

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using glass beads vortexing. Includes lysis buffer recipes. The Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using glass beads vortexing]

Preparation of Crude Subcellular Fractions by Differential Centrifugation Protocol - http://www.axis-shield.com/densityhome/optiprep/S07.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

The employment of differential centrifugation to prepare crude fractions of subcellular particles from homogenates is often a necessary first step to a subsequent purification of one or more particles on a density gradient. This protocol describes the use of differential centrifugation to fractionate a mammalian liver homogenate but similar methods should be applicable to all mammalian tissues and cultured cells. - [Read Preparation of Crude Subcellular Fractions by Differential Centrifugation Protocol]

Preparation of RNA from Plant Tissue Using Trizol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4105

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol is based on the standard Trizol protocol for the purification of RNA from animal cells using Trizol (Purification of RNA from Animal Cells using Trizol). In this version, adapted for use with plant tissues, a high-salt isopropanol precipitation step has been added to precipitate RNA selectively, while maintaining polysaccharides and proteoglycans in solution. - [Read Preparation of RNA from Plant Tissue Using Trizol]

Production of dsRNA for RNAi in Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4503

Category: Avian Bird Protocols

Protocol describes the production of double-stranded RNA (dsRNA) from fragments of cDNAs of candidate genes. The cDNA fragments must be cloned in plasmids with a flanking SP6 and T7 promoter (e.g., pSP72 or pCRII). The plasmid is linearized and sense and antisense RNAs are produced separately by in vitro transcription. After purification, the RNA strands are annealed to yield a dsRNA molecule suitable for RNAi in avian embryos. - [Read Production of dsRNA for RNAi in Avian Embryos Protocol]

Protein Expression and Purification Flowchart - http://www.embl.de/ExternalInfo/protein_unit/draft_frames/frame_flowchart_ext.htm

Category: Protein Protocols: Protein Expression Protocols

Protein Expression and Purification Flowchart. EMBL - [Read Protein Expression and Purification Flowchart]

Protein Expression and Purification Protocol - http://sosnick.uchicago.edu/Protein_Express&Purify.html

Category: Recombinant Protein Protocols

From plasmid to protein using bacterial expression. Transform appropriate DNA plasmid, Make a starter culture for protein expression, bacteria culture for protein expression. Sosnick Group Chicago. - [Read Protein Expression and Purification Protocol]

Protein Precipitation Protocols - http://www.ls.huji.ac.il/~purification/Protocols/ProteinPrecipitation.html

Category: Protein Protocols: Protein Precipitation Procols

TCA-DOC Normal, TCA Acetone Precipitation, Ethanol Precipitation, TCA-DOC / Acetone, Acidified Acetone/Methanol, Dr. Mario Lebendiker. University of Jerusalem. - [Read Protein Precipitation Protocols]

Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole - http://www.cshprotocols.org/cgi/content/extract/2006/9/pdb.prot4262

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Perform this protocol after determining the imidazole concentration that yields the best purification of the recombinant protein of interest. - [Read Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole]

Protein Purification: Assays, Specific Activity, Initial Fractionation - http://wine1.sb.fsu.edu/bch5425/lect29/lect29.htm

Category: Protein Protocols: Protein Purification and Identification

Protein Purification: Assays, Specific Activity, Initial Fractionation. Blaber Lecture. - [Read Protein Purification: Assays, Specific Activity, Initial Fractionation]

Protein Purification: Column Chromatography - Ion exchange; Dialysis and Concentration - http://wine1.sb.fsu.edu/bch5425/lect30/lect30.htm

Category: Protein Protocols: Protein Purification and Identification

Protein Purification: Column Chromatography - Ion exchange; Dialysis and Concentration. Blaber lecture - [Read Protein Purification: Column Chromatography - Ion exchange; Dialysis and Concentration]

Protein Purification: Gel Filtration, Affinity and Hydrophobic resins; Preparation of Resin, Plumbin - http://wine1.sb.fsu.edu/bch5425/lect31/lect31.htm

Category: Protein Protocols: Protein Purification and Identification

Protein Purification: Gel Filtration, Affinity and Hydrophobic resins; Preparation of Resin, Plumbing. Blaber Lecture. - [Read Protein Purification: Gel Filtration, Affinity and Hydrophobic resins; Preparation of Resin, Plumbin]

Protocol for Immunopurification of Tyrosine Phosphorylated Proteins - http://www.bdbiosciences.com/pharmingen/protocols/Tyrosine_Phosphorylated_Proteins.shtml

Category: Signalling Signal Transduction Protocols

Protocol for immunopurification of tyrosine phosphorylated proteins. Protocol includes: Cell Lysis (Non-Denaturing and Denaturing Conditions) and Purification. - [Read Protocol for Immunopurification of Tyrosine Phosphorylated Proteins]

Protocol for the Isolation of a Heterogenous Muscle-Derived Cell Population - http://www.bcm.edu/labs/goodell/protocols/Goodell_Musclestemcells.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol is employed for the purification of a population of muscle-derived cells from skeletal muscle of C57Bl/6 animals. The resulting preparation is a mixture of many cell types including satellite cells (a.k.a., muscle progenitor cells) and hematopoietically active muscle-derived cells. - [Read Protocol for the Isolation of a Heterogenous Muscle-Derived Cell Population]

Protocol: Purification of In Vitro Synthesized mRNA with Microcon or Centricon Centrifugal Filters - http://www.millipore.com/techpublications/tech1/6dlk4v

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

In vitro transcription reactions employing T3, T7 or SP6 phage-encoded RNA polymerases are widely used to synthesize RNA from recombinant vectors containing appropriate promoters. Production of large amounts of specific RNA is valuable in the preparation of hybridization probes and in vitro translation studies; in the synthesis of ribozymes, rRNA, SRP, antisense RNA and substrates for RNA splicing; and in RNA-protein interaction studies. - [Read Protocol: Purification of In Vitro Synthesized mRNA with Microcon or Centricon Centrifugal Filters]

Purification by affinity chromatography of H1Â° RNA-binding - http://147.52.72.117/IJMM/2003/volume11/number4/509.pdf

Category: Chromatography Protocols

Purification by affinity chromatography of H1Â° RNA-binding proteins from rat brain. MARIA SCATURRO et al. - [Read Purification by affinity chromatography of H1Â° RNA-binding]

Purification Of Anti Peptide Antibody - Claire Walczak - http://mitchison.med.harvard.edu/protocols/ab4.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Purification Of Anti Peptide Antibody - Claire Walczak - [Read Purification Of Anti Peptide Antibody - Claire Walczak]

Purification of Antibodies on an Antigen Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4284

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Immunoaffinity purification of antibodies is used to purify antigen-specific antibodies from a preparation of polyclonal antibodies. Such purification is commonly needed in the production of antipeptide antibodies, where it is used to concentrate the desired antibodies and separate them from those raised against carrier proteins. It is also used for the more general purpose of removing unwanted, nonspecific binding activity from polyclonal antibody preparations. - [Read Purification of Antibodies on an Antigen Column Protocol]

Purification of BAC DNA Prepared with the NucleoBond System Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4391

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol describes purification of BAC DNA that has been prepared with the commercial NucleoBond system. - [Read Purification of BAC DNA Prepared with the NucleoBond System Protocol]

Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3990

Category: Viral Protocols

Protocol is used to prepare the arms of any bacteriophage {lambda} vector. - [Read Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol]

Purification of Bacteriophage lambda Particles by Isopycnic Centrifugation through CsCl Gradients - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3968

Category: Viral Protocols

Isopycnic centrifugation through CsCl gradients is used to prepare infectious bacteriophage {lambda} particles of the highest purity that are essentially free of contaminating bacterial nucleic acids. - [Read Purification of Bacteriophage lambda Particles by Isopycnic Centrifugation through CsCl Gradients]

Purification of Bovine Lens and Bacterially Expressed Human Vimentin Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4675

Category: Cytoskeleton Protocols: Intermediate Filaments Protocols

Intermediate filaments (IF) are major cytoskeletal systems of vertebrate and many nonvertebrate cells whose expression is cell-type specific and developmentally regulated. This protocol describes a method for purifying one type of IF, vimentin, from bovine lens tissue. Purification of human vimentin expressed in Escherichia coli is also described. These methods are useful in the preparation of other IF protein subunits for microinjection studies as well. - [Read Purification of Bovine Lens and Bacterially Expressed Human Vimentin Protocol]

Purification of Caveolae Membranes from a Plasma Membrane Fraction of Cultured Cells and Tissues - http://www.axis-shield.com/densityhome/optiprep/S19.pdf

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

The first part of the isolation procedure is a flotation through a continuous iodixanol gradient; this gradient is essentially a resolving gradient in which the caveolin-rich vesicles are concentrated in the top third of the gradient, while the predominantly caveolin-poor vesicles band in denser regions. A second discontinuous gradient is essentially a concentration gradient to band the caveolin-rich vesicles sharply at an interface. - [Read Purification of Caveolae Membranes from a Plasma Membrane Fraction of Cultured Cells and Tissues]

Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradient - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3913

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

A solution containing plasmid DNA, saturating amounts of ethidium bromide, and CsCl (44% w/v) is layered between two solutions of lesser (35% w/v CsCl) and greater density (59% w/v CsCl). During centrifugation to equilibrium, the closed circular plasmid DNA and linear DNAs form bands at different densities. - [Read Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradient]

Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradients - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3927

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Solutions containing plasmid DNA are adjusted to a density of 1.55 g/ml with solid CsCl. The intercalating dye, ethidium bromide, which binds differentially to closed circular and linear DNAs, is then added to a concentration of 200 mu;g/ml. During centrifugation to equilibrium, the closed circular DNA and linear DNAs form bands at different densities. - [Read Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradients]

Purification of Demethylated Sphingomyelin Protocol - http://www.duke.edu/web/ceramide/protocols/0032.html

Category: Lipid Protocols

Protocol for the purificiation of demethylated sphingomyelin. Includes: Lower, chloroform phase; Upper, aqueous phase and interphase. - [Read Purification of Demethylated Sphingomyelin Protocol]

Articles (1-4 of 4)

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.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

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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