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Purification of DNA Recovered Anion-exchange Chromatography Protocol - http://cshprotocols.cshlp.org/cgi/content/extract/2006/2/pdb.prot4024

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for the purification of DNA recovered from agarose and polyacrylamide gels by anion-exchange chromatography. Fragments of DNA recovered from agarose gels are sometimes poor templates or substrates in subsequent enzymatic reactions. This problem can be solved by binding the DNA to a positively charged matrix, such as DEAE-Sephadex or DEAE-Sephacel, in buffers of low ionic strength. After washing the matrix, the DNA is eluted by raising the strength of the buffer. - [Read Purification of DNA Recovered Anion-exchange Chromatography Protocol]

Purification of Fusion Proteins by Affinity Chromatography on Glutathione Agarose Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3655

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant proteins, constructed in pGEX vectors, are fused to glutathione S-transferase (GST) and can be purified to near homogeneity by affinity chromatography on glutathione-agarose. Bound GST-fusion proteins are readily displaced from the column by elution with buffers containing free glutathione. - [Read Purification of Fusion Proteins by Affinity Chromatography on Glutathione Agarose Protocol]

Purification of Golgi Membranes from a Light Mitochondrial Fraction in a Self-Generated Gradient - http://www.axis-shield.com/densityhome/optiprep/S12.pdf

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

This protocol uses a "light mitochondrial" pellet from a mammalian liver homogenate. The gradient thus has to resolve a variety of denser components (peroxisomes, lysosomes, mitochondria) from the Golgi membranes, which have a low density in iodixanol (1.06-1.09 g/ml) [1]. The protocol is specifically tailored to the purification of Golgi membranes from this pellet and is unsuitable for the isolation or analysis of other organelles present in the light mitochondrial fraction. - [Read Purification of Golgi Membranes from a Light Mitochondrial Fraction in a Self-Generated Gradient]

Purification of GST-fusion proteins E. coli. (pGEX vector) Protocol - http://mullinslab.ucsf.edu/protocols/html/GST_fusion_expression.htm

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the purification of GST-fusion proteins E. coli. (pGEX vector). - [Read Purification of GST-fusion proteins E. coli. (pGEX vector) Protocol]

Purification of Histidine-tagged Proteins by Immobilized Ni2+ Absorption Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4088

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant proteins engineered to have a polyhistidine tail at either the carboxyl or amino terminus can easily be purified in one step by affinity chromatography on a resin carrying chelated nickel ions. Chromatography can be carried out in column or batch formats. After unbound proteins are washed away, the target protein is eluted using imidazole, which typically preserves the antigenic and functional features of the protein. - [Read Purification of Histidine-tagged Proteins by Immobilized Ni2+ Absorption Chromatography Protocol]

Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4221

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Immobilized metal-ion affinity chromatography (IMAC) is suitable for the purification of proteins under denaturing conditions. Either guanidine-HCl or urea can be used, although guanidine-HCl is a stronger denaturant than urea. Proteins that have been adsorbed to the column in the presence of guanidine-binding buffer may be washed with urea-binding buffer and eluted with urea elution buffer. - [Read Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol]

Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4217

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Histidine-tagged proteins can be purified on prepacked 1-ml immobilized metal-ion affinity chromatography (IMAC) columns without optimization of the separation conditions. The method allows fast capture of the target protein, although with a lower purity than can be obtained under optimized conditions. - [Read Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol]

Purification of Human Mononuclear Cells and Neutrophils Protocol - http://www.cbrinstitute.org/labs/springer/protocols/chunduk_mono_neutro.html

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for the purification of human mononuclear cells and neutrophils. - [Read Purification of Human Mononuclear Cells and Neutrophils Protocol]

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

Category: Nucleic Acid Purification Protocols

Protocol for purification of in vitro synthesized mRNA with Microcon or Centricon Centrifugal Filters. - [Read Purification of In Vitro Synthesized mRNA with Microcon or Centricon Centrifugal Filters Protocol]

Purification of Labeled Probes Using the High Pure PCR Product Purification Kit Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_71-72.pdf

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Protocol for purification of labeled probes using the High Pure PCR Product Purification Kit. - [Read Purification of Labeled Probes Using the High Pure PCR Product Purification Kit Protocol]

Purification of Labeled Probes using the High Pure PCR Product Purification Kit Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_71-72.pdf

Category: Nucleic Acid Purification Protocols

Protocol for purification of labeled probes using the High Pure PCR Product Purification Kit. - [Read Purification of Labeled Probes using the High Pure PCR Product Purification Kit Protocol]

Purification of leukocyte fractions from rat peritoneal exudates by density barrier sedimentation. - http://www.axis-shield.com/densityhome/optiprep/C19.pdf

Category: Cell Biology and Cell Culture

The separation depends on the use of a hyperosmotic density barrier to separate the two types of leukocyte. - [Read Purification of leukocyte fractions from rat peritoneal exudates by density barrier sedimentation.]

Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio - http://www.fhcrc.org/science/labs/fero/Protocols/Hematopoietic.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Antibodies for Lineage Depletion; Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio]

Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells Protocol - http://www.fhcrc.org/science/labs/fero/Protocols/Hematopoietic.pdf

Category: Immunology

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells Protocol]

Purification of Maltose-binding Fusion Proteins by Affinity Chromatography on Amylose Resin Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4087

Category: Chromatography Protocols: Affinity Chromatography Protocols

Foreign proteins fused to maltose-binding protein can be readily purified to near homogeneity by affinity chromatography on resins containing cross-linked polysaccharides such as amylose. - [Read Purification of Maltose-binding Fusion Proteins by Affinity Chromatography on Amylose Resin Protocol]

Purification of Mammalian Liver Mitochondria by Flotation Through a Pre-formed Discontinuous Iodixan - http://www.axis-shield.com/densityhome/optiprep/S15.pdf

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

This protocol describes a discontinuous gradient, which resolves the mitochondria from both lighter and denser organelles. Because the centrifugation is carried out for 4 h, diffusion will create a partially continuous gradient and this probably contributes to the resolution of the mitochondria from the lighter lysosomes. - [Read Purification of Mammalian Liver Mitochondria by Flotation Through a Pre-formed Discontinuous Iodixan]

Purification of Monocytes Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protocols.html~main

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for the purification of monocytes. - [Read Purification of Monocytes Protocol]

Purification of Neutrophils Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protocol1.html~main

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for purification of neutrophils. - [Read Purification of Neutrophils Protocol]

Purification of Nuclear Factor DNA Recognition Site Affinity PDF - http://www.jbc.org/cgi/reprint/261/3/1398.pdf

Category: Chromatography Protocols

Purification of Nuclear Factor DNA Recognition Site Affinity. PDF. - [Read Purification of Nuclear Factor DNA Recognition Site Affinity PDF]

Purification of Nucleic Acids by Extraction with Phenol:Chloroform Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4455

Category: Nucleic Acid Purification Protocols

Protocol describes the standard method for nucleic acid purification by extraction first with phenol:chloroform (optionally containing hydroxyquiniline at 0.1%) and then with chloroform to remove any remaining phenol. The procedure takes advantage of the fact that deproteinization is more efficient when two different organic solvents are used instead of one. - [Read Purification of Nucleic Acids by Extraction with Phenol:Chloroform Protocol]

Purification of Oocyst Walls and Sporocysts from Toxoplasma gondii Protocol - http://www.axis-shield.com/densityhome/optiprep/C34.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Although Percoll gradients were able to provide a purified sporocyst fraction, because these particles do not all band in a discrete manner in such gradients, they were unable to provide a simultaneous isolation of a pure oocyst wall fraction. Gradients formed from this protocol on the other hand are able to provide purified sporocysts and oocyst walls in the same gradient. - [Read Purification of Oocyst Walls and Sporocysts from Toxoplasma gondii Protocol]

Purification of PCR Products in Preparation for Cloning Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3825

Category: Cloning Protocols

Protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning. - [Read Purification of PCR Products in Preparation for Cloning Protocol]

Purification of PCR Products in Preparation for Cloning Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3825

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning. - [Read Purification of PCR Products in Preparation for Cloning Protocol]

Purification of Peptides from Solid-Phase Peptide Synthesis with RP-HPLC Protocol - http://cshprotocols.cshlp.org/cgi/content/abstract/2006/28/pdb.prot4548

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol details the purification and analysis of many synthetic peptides of 2-65 amino acid residues. These peptides contain a number of ionizable or polar side chains, but do not contain secondary structural elements (such as ß-sheets) that favor supramolecular assembly. - [Read Purification of Peptides from Solid-Phase Peptide Synthesis with RP-HPLC Protocol]

Purification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation - http://www.plantphysiol.org/cgi/reprint/75/3/670

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

A procedure developed to purify simultaneously peroxisomes and mitochondria from spinach (Spirnacia olercea L.) leaf under isoosmotic and low viscosity conditions. This method involves differential centrifugation and density gradient centrifugation on four layers of Percoll. - [Read Purification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation]

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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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