solid Protocols

Category: Cell Culture Protocols: Cell Staining Protocols

A simple method for handling suspension cells is to attach them to a solid substrate before fixation, and one way to do this is to use a cytocentrifuge. - [Read Attaching Suspension Cells to Slides Using a Cytocentrifuge Protocol]

Category: Antibody Protocols: Epitope Mapping Antibody Protocols

A set of overlapping synthetic peptides is synthesized, each corresponding to a small segment of the linear sequence of a protein antigen and arrayed on a solid phase. The panel of solid-phase peptides is then probed with a test antibody, and bound antibody is detected using an enzyme-labeled secondary antibody. This method is very rapid and can be extraordinarily successful. - [Read Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

The most commonly used markers for selection of transgenic Arabidopsis are resistance to the antibiotic kanamycin and to the herbicide glufosinate ammonium. Resistance to kanamycin is conferred by a bacterial gene encoding the enzyme neomycin phosphotransferase (NPT). In this protocol, kanamycin-resistant seedlings are selected on solid medium. - [Read Kanamycin Selection of Transformed Arabidopsis Protocol]

Category: Yeast Protocols: Yeast Transformation Protocols

Protocol for large-scale yeast transformation. Includes: Yeast Cell Preparation; Large Scale Transformation; To plate on solid medium; To select in liquid. - [Read Large-Scale Yeast Transformation Protocol]

Category: DNA Microarray Protocols

Making a scanning array: in situ synthesis of oligonucleotides on a solid substrate. Elder et al., Oxford Practical Approach Series. - [Read Making a scanning array: in situ synthesis of oligonucleotides on a solid substrate]

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

The yeast, Saccharyomyces cerevisiae, has become an important organism in molecular, biochemical, and genetic analysis. The organism has specific requirements for growth under a variety of conditions. The media, both liquid and solid, simple, define, and complex are describe in this unit. Also included are methods for handling, storing, and shipping stock of yeast. - [Read Media and Culture of Yeast Protocol]

Category: Microsphere Analysis Protocols

The role of microspheres in these screens is similar to their traditional role in immunoassays, namely as a solid phase to either enhance detection, separation, or both. The predominance of radioactive assays in high-throughput screening, along with the desire to find alternative means of detection, have led to research on substituting alternative fluorescent technologies. - [Read Microspheres for High-Throughput Screening Assays]

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

This protocol is the first of two describing solid-phase chemical cleavage of mismatch (SpCCM), a method for the detection of mutations in genomic DNA. DNA fragments up to 2 kb in length can be analyzed to determine the position (within 10-15 bp) and identity of the mismatched nucleotide. - [Read Mutation Detection by Solid-Phase Chemical Cleavage of Mismatches Using Radioactive Probes Protocol]

Category: Fungi Protocols

Protocol for the preparation of solid tissue for Aspergillus galactomannan antigen detection by Platelia (Biorad). Technique was designed for use on human serum. However, it may also be possible to perform this method on solid tissues and organic solutions. Viscous solution and tissue specimens need to be pre-treated to achieve the extraction of the Aspergillus antigen and to get a homogeneous sample in solution. - [Read Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA in solid tissue samples using either propidium iodide (PI) or DAPI staining followed by flow cytometry. PI can be excited at 488 nm by the argon-ion laser, the most commonly used laser in flow cytometry. Alternatively, DAPI is best excited by a high-power UV laser, which is less commonly available. - [Read Propidium Iodide (PI) or DAPI Staining of Unfixed Solid Tissues for Flow Cytometry Protocol]

Category: Peptide Protocols

Protocol for Manual Peptide Synthesis. Resin Swelling And Coupling Of Activated Amino Acid Esters. Deblocking and cleaving peptide from solid support. Synthesis of Peptides. The University of Nebraska-Lincoln Protein Core Facility - [Read Protocol for Manual Peptide Synthesis]

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]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Protocol describes a method for storing bacterial cultures that are growing on solid media. It is useful for storing copies of cDNA. - [Read Storage of Bacterial Cultures Growing on Solid Medium Protocol]