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Alkaline Agarose Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4027

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

Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase) and to analyze the size of the DNA strand after digestion of DNA-RNA hybrids with nucleases such as S1. - [Read Alkaline Agarose Gel Electrophoresis Protocol]

Analysis of Recombinant Bacteriophage M13 Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3996

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A rapid method to analyze the size of the single-stranded DNA of M13 recombinants. - [Read Analysis of Recombinant Bacteriophage M13 Clones Protocol]

Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4426

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing ES cell-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for ES cell-diploid embryo aggregation in which diploid embryos are replaced with tetraploid embryos. The resulting chimeras can be used to analyze the embryonic versus extraembryonic phenotype of a mutation. - [Read Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Immunology: B Cell Protocols

Fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4560

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Protocol describes the use of chromatin immunoprecipitation technology (ChIP) to analyze interactions of proteins or protein complexes with DNA in vivo. In this approach, the material is fixed with formaldehyde to preserve DNA-protein and protein-protein associations, the cells are lysed, and the chromatin is cut and solubilized. The chromatin suspension is immunoprecipitated with an antibody against the protein(s) of interest, and the coimmunoprecipitated DNA fragments are analyzed. - [Read Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol]

Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana - http://vkp.leeds.ac.uk/Drive/Drive/ViewFile?include=y&component=security&action=Start&nextAction=gotoDoc&doc=494824#search=%22chromatin%20immunoprecipitation%20protocol%22

Category: Epigenetics and Methylation Protocols: ChIP Chromatin Immunoprecipitation Protocols

Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana. Werner Aufsatz, Matzke Lab. Gregor Mendel Institute of Molecular Plant Biology, Austria. Includes Chromatin Crosslinking, Chromatin preparation, Pre-clearing and immuno precipitation (IP), Collection,washes and elution of immune complexes, Reverse crosslinking and DNA cleanup - [Read Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells. Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: Western Blot Protocols

Extraction of Total Protein from Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4680

Category: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

The simplest way to analyze proteins is in unfractionated extracts. However, it is often desirable to fractionate proteins, e.g, by size. This procedure extracts total protein from Arabidopsis samples. Typical yields are ~2-3 mg/ml (using rosette leaves) or 6-8 mg/ml (using young seedlings). - [Read Extraction of Total Protein from Arabidopsis Protocol]

In Vitro System to Analyze Light Signal Transduction in Neurospora crassa - http://www.fgsc.net/neurosporaprotocols/How%20to%20analyze%20light%20signal%20transduction%20in%20vitro.pdf

Category: Fungi Protocols: Neurospora Protocols

Procedure for an in vitro system to analyze light signal transduction in Neurospora crassa - [Read In Vitro System to Analyze Light Signal Transduction in Neurospora crassa]

Method to Analyze Meiotic Segregation Patterns in Coprinus cinereus Using PCR - http://www.fgsc.net/fgn/freedman.html

Category: Fungi Protocols: Fungi Genetics Protocols

Quick and reliable method to analyze meiotic segregation patterns in Coprinus cinereus using the polymerase chain reaction. The advantages of this method include: 1. The tissue is grown and lyophilized in the same tube, which facilitates the simultaneous analysis of many segregants. 2. Only one extraction step is necessary. 3. The markers are scored by gel electrophoresis, thereby bypassing Southern analysis. - [Read Method to Analyze Meiotic Segregation Patterns in Coprinus cinereus Using PCR]

Preparation of Yeast Cells for Flow Cytometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4178

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Flow cytometry is used to analyze the quantity of DNA in cells. Since the DNA content of cells varies through the cell cycle, this information can provide an indication of cell cycle progression. This protocol uses SYTOX Green staining. - [Read Preparation of Yeast Cells for Flow Cytometry Protocol]

Synthesis of DNA Libraries: Use of PCR to Analyze Ligation Substrates and Products - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4136

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

A vector that lacks inserts can significantly contaminate a DNA library. This contamination can occur either from self-ligation of single-cut vector or from uncut circular starting plasmid. - [Read Synthesis of DNA Libraries: Use of PCR to Analyze Ligation Substrates and Products]

Use of PCR for Quality Control of a Genomic DNA Library Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4138

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol describes the use of PCR to analyze the quality of a human genomic DNA library. It has been optimized for a human genomic DNA library containing fragments between approximately 50 and 500 bp. - [Read Use of PCR for Quality Control of a Genomic DNA Library Protocol]

Articles (1-3 of 3)

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.