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(LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4107

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

LCM isolates specific cells or tissues from samples mounted on microscope slides. The samples are viewed through a thermoplastic film that is attached to a microcentrifuge tube lid. Localized heat, caused by the application of a laser pulse, fuses the membrane to the cells of interest, which can then be harvested for further analysis. RNA and proteins can be purified from the isolated cells, allowing detailed analysis of gene expression. This protocol is divided into three stages. - [Read (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel]

A non-radioactive electrophoretic mobility shift assay for the detection of heat shock element (HSE) - http://www.fgsc.net/fgn45/45meyer.html

Category: DNA Protocols: EMSA DNA-Protein Protocols

A non-radioactive electrophoretic mobility shift assay based on the digoxigenin detection system had been developed and applied to the analysis of the interaction between the heat shock transcription factor and the heat shock element of N. crassa. (Bioch - [Read A non-radioactive electrophoretic mobility shift assay for the detection of heat shock element (HSE)]

How to Test Heat, Osmotic, and Oxidative Tolerance of Neurospora Conidia and Young Mycelia - http://www.fgsc.net/neurosporaprotocols/How%20to%20test%20thermal%20osmotic%20and%20oxidative%20stress%20tolerance.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to test heat, osmotic, and oxidative tolerance of Neurospora conidia and young mycelia. - [Read How to Test Heat, Osmotic, and Oxidative Tolerance of Neurospora Conidia and Young Mycelia]

Northern Hybridization Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3723

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol for northern hybridization. Protocol describes how to carry out northern hybridization at high stringency in phosphate-SDS-buffers. Although a wide variety of formats are available, hybridization is usually performed in heat-sealable bags, roller bottles, or plastic boxes, as described here. - [Read Northern Hybridization Protocol]

Protocol for Immunofluorescence - http://www.bio.unc.edu/faculty/salmon/lab/protocolsimmunofluorescence.html

Category: Molecular Imaging: Immunofluorescence Microscopy

Protocol for Immunofluorescence - Salmon Lab. Making Boiled Donkey (or whatever) Serum (BDS) for Blocking. Making Heat-Inactivated Serum for Blocking. Mounting Media. - [Read Protocol for Immunofluorescence]

Single-Strand Conformation Polymorphism Analysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4118

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Single-strand confirmation polymorphism analysis (SSCP) is a powerful and robust method for the detection of DNA sequence changes (single-base substitutions) based on shifts in electrophoretic mobility. In this protocol, the target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Single-Strand Conformation Polymorphism Analysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4118

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

The target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. Differences in sequence alter the conformation of the DNA and hence its electrophoretic mobility and, because of the high resolution of polyacrylamide gels, most conformational changes caused by subtle changes in sequence can be detected. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4044

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol for southern hybridization of radiolabeled probes to nucleic acids immobilized on membranes. Protocol describes how to carry out Southern hybridizations at high stringency in phosphate-SDS buffers. Although a wide variety of formats are available, most Southern hybridizations are carried out in heat-sealable bags, roller bottles, or plastic boxes. - [Read Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes Protocol]

Unmasking Hidden Epitopes Using the Microwave Oven Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4529

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Several methods have been developed to "retrieve" antigens that have been masked by fixation. The principle behind using the microwave oven method described here is to use extended periods of heat to break some of the subcellular structures that block antibody access. Be aware that any of the antigen retrieval methods should be avoided wherever possible, because they may introduce artifactual false-positive staining. - [Read Unmasking Hidden Epitopes Using the Microwave Oven Protocol]

Unmasking Hidden Epitopes Using the Pressure Cooker Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4530

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

The principle behind the pressure cooker method described here is to use extended periods of heat to break some of the subcellular structures that block antibody access. This approach is appropriate for handling specimens on glass slides. The major advantages of the pressure cooker method are the ability to handle a large number of slides simultaneously, the convenience of using metal racks, and the avoidance of any hot spots that are found in the microwave. - [Read Unmasking Hidden Epitopes Using the Pressure Cooker Protocol]

Articles (1-7 of 7)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

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.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

Microinjection Pipette Pulling Protocol

A simple protocol for microinjection pipette pulling.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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