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Configuration Column Construction Column Packing for Capillary Liquid Chromatography - http://cshprotocols.cshlp.org/cgi/content/abstract/2007/3/pdb.prot4578

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for configuration, column construction, and column packing for a capillary liquid chromatography system. Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to several commercial UV detectors. - [Read Configuration Column Construction Column Packing for Capillary Liquid Chromatography]

Construction of Micro-Tip for Use in IMAC Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000163.pdf?pid=PP00000163

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the second step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LCMS/ MS. This procedure describes the construction of microchromatographic columns, or micro-tips. - [Read Construction of Micro-Tip for Use in IMAC Protocol]

DNA Extraction from Agarose Gels Protocol - http://www.methodbook.net/dna/gelextrc.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

DNA Extraction from Agarose Gels Protocol. The page includes cutting out the DNA band from the gel, and describes three methods including 1) Spin-columns (Nucleic acid purification columns), 2) using Dialysis tubing (semi-permeable membrane, Visking tubing), and the 3) Paper strip method.Matt Lewis, Department of Pathology, University of Liverpool. - [Read DNA Extraction from Agarose Gels Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: BACs Bacterial Artificial Chromosome Protocols

Rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

FPLC Protocol - http://users.path.ox.ac.uk/~ciu/Methods/Protein/fplc.html

Category: Chromatography Protocols

FPLC Protocol. The FPLC consists of a pump and a column which will withstand high pressure so separations can be carried out relatively quickly. For a detailed description there is a FPLC system handbook which is particularly useful for trouble shooting. For use of individual columns follow the "instructions" (in the green folder) which accompany each one. Sir William Dunn School of Pathology, Oxford University. - [Read FPLC Protocol]

Fragment purification on Sephacryl S-500 spin columns - http://www.genome.ou.edu/protocol_book/protocol_partI.html#I.H

Category: Chromatography Protocols: Column Chromatography Protocols

Fragment purification on Sephacryl S-500 spin columns , Bruce A. Roe. - [Read Fragment purification on Sephacryl S-500 spin columns]

HPLC Column Care Protocols and Information - http://www.nestgrp.com/pdf/colcare.pdf

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocols and information on general HPLC column care. Includes: Silica based columns and Polymer based columns. Protocols included: pH stability, Mechanical stability, Mobile phases (Eluents), Proper storage of HPLC columns, Equilibration time, Regeneration of a column, Regeneration of RP packings, Regeneration of NP (Normal Phase) packings, Regeneration of Ion Exchange Packings - [Read HPLC Column Care Protocols and Information]

Immunoaffinity Purification: Coupling Antibodies to Protein A or G Bead Columns Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4303

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes how antibodies are bound to protein A or protein G beads and are then crosslinked to the matrix via a bifunctional coupling reagent called dimethyl pimelimidate (DMP). - [Read Immunoaffinity Purification: Coupling Antibodies to Protein A or G Bead Columns Protocol]

Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4276

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

This elution procedure relies on disrupting the bonds between the antibody and the antigen. - [Read Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns Protocol]

Nonradioactive Cycle Sequencing of PCR-Amplified DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4095

Category: DNA Protocols: DNA Sequencing Protocols: Big Dye Protocols

Protocol describes the purification, quantification, andsubsequent sequencing of amplified DNA fragments using PCR.Excess nucleotides are removed from the initial PCR productsusing spun columns, and the products are quantified using fluorometry. - [Read Nonradioactive Cycle Sequencing of PCR-Amplified DNA Protocol]

Packing Capillary Columns for RP-HPLC Protocol - http://cshprotocols.cshlp.org/cgi/content/abstract/2006/28/pdb.prot4546

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to a number of commercial UV detectors. - [Read Packing Capillary Columns for RP-HPLC Protocol]

Packing Ceramic Hydroxyapatite Columns for High-Performance Liquid Chromatography Protocol - http://cshprotocols.cshlp.org/cgi/content/extract/2006/9/pdb.prot4267

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes the packing of ceramic hydroxyapatite for HPLC. - [Read Packing Ceramic Hydroxyapatite Columns for High-Performance Liquid Chromatography 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 Synthetic Oligonucleotides by Polyacrylamide Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4063

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Polyacrylamide Gels Protocols

Protocol for purification of synthetic oligonucleotides by polyacrylamide gel electrophoresis. As a rule of thumb, oligonucleotides >25 nucleotides should be purified by polyacrylamide gel electrophoresis, as should oligonucleotides of any length that yield anomalous results. After electrophoresis, the oligonucleotide is eluted from the gel and concentrated by reversed-phase chromatography on Sep-Pak C18 columns. - [Read Purification of Synthetic Oligonucleotides by Polyacrylamide Gel Electrophoresis Protocol]

Rapid Elution of DNA Agarose Gels Protocol - http://www.fhcrc.org/science/labs/hahn/methods/mol_bio_meth/dna_spin_ext.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

Rapid Elution of DNA Agarose Gels Protocol. This method allows quick purification of DNA fragments from agarose gels for use in cloning and other reactions. Higher yields of purified DNA can be obtained from commercially available purification kits, however greater ligation efficiencies per given amount of DNA have been seen with the use of these described spin columns. Hahn Lab. - [Read Rapid Elution of DNA Agarose Gels Protocol]

Selection of Poly(A)+ RNA by Oligo(dT)-Cellulose Chromatography - Subscription Required - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4047

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

Chromatography on oligo(dT) columns is the preferred method for large-scale purification (>25 Âµg) of poly(A)+ RNA extracted from mammalian cells. Typically, between 1% and 10% of the RNA applied to the oligo(dT) column is recovered as poly(A)+ RNA. Because the method can be frustratingly slow, it is not recommended for purification of poly(A)+ RNA from multiple samples. For this purpose, batch elution (Selection of Poly(A)+ RNA by Batch Chromatography) is the better choice. - [Read Selection of Poly(A)+ RNA by Oligo(dT)-Cellulose Chromatography - Subscription Required]