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Fixing Attached Cells in Paraformaldehyde Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4294

Category: Cell Culture Protocols: Cell Fixing Protocols

Treating cells with paraformaldehyde leads to the establishment of chemical cross-links between free amino groups. When the cross-links join different molecules, a latticework of interactions occurs that holds the overall architecture of the cell together. Commercial formaldehyde solutions are not recommended, because they lack the advantages of using a variable-length polymer, and the cells will simultaneously be fixed with the alcohol (usually methanol). - [Read Fixing Attached Cells in Paraformaldehyde Protocol]

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]

Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4207

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol describes a general approach for the direct attachment of a triazine dye onto a carbohydrate support. The direct coupling method is achieved under alkaline conditions (pH 10-11) with nucleophilic displacement of the dye’s chlorine atom by the polymer hydroxyls. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol]

Recovery of DNA from Low-melting-temperature Agarose Gels: Enzymatic Digestion with Agarase Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4026

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

A fragment of gel containing a band of DNA is excised and digested with agarase, which hydrolyzes the polymer to disaccharide subunits. The released DNA is then purified by phenol extraction and ethanol precipitation. The method works well for DNAs ranging in size from <5 kb to >20 kb. - [Read Recovery of DNA from Low-melting-temperature Agarose Gels: Enzymatic Digestion with Agarase Protocol]

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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.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

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