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Measurement of Intercellular Conjugates by Flow Cytometry Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C6BB1DCEB151C1340A080226633&objectid=6674F2EBA1D257D06F0E2E6B88848643

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

The more commonly available single-laser cytometers can also be used to measure multicellular conjugates, but due to overlaps in emission spectra, the extent of labeling cells with fluorophores must be controlled much more carefully when the single-laser machines are used. This protocol describes the labeling of cells and analysis of conjugates with either dual-laser or single laser flow cytometers. - [Read Measurement of Intercellular Conjugates by Flow Cytometry Protocol]

Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4255

Category: Western Blot Protocols: Antigen Detection Methods Protocols

Protocol describes, samples containing the target protein are deposited onto a polyvinyldifluoride (PVDF) membrane using a vacuum manifold. The immobilized protein is exposed to an antibody specific for the target protein, followed by an antibody that reacts with species-specific determinants carried by the primary antibody and is conjugated to horseradish peroxidase (HRP). - [Read Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection]

Media and Culture of Yeast Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E5192D47CE9B0776DB7D136C34ED148A&objectid=6673ABA6C2DB2E4E7564EDFA26589256

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]

Membrane Potential Analysis by Flow Cytometry Protoocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/membrane.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for membrane potential analysis by flow cytometry. Includes: Preparation of the dyes; Loading of the dyes; Flow Cytometric Analysis. - [Read Membrane Potential Analysis by Flow Cytometry Protoocol]

Metabolic Labeling of Plant Cell Cultures with K15NO3 Protocol - http://www.plantmethods.com/content/2/1/14

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Protocol for metabolic labeling of plant cell cultures with K15NO3 as a tool for quantitative analysis of proteins and metabolites. - [Read Metabolic Labeling of Plant Cell Cultures with K15NO3 Protocol]

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]

Methods for Plant RNA Isolation - http://wheat.pw.usda.gov/~lazo/methods/lazo/met6.html

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

AMES/Chloroform extraction to extract total RNA from potato leaves for viroid analysis. - [Read Methods for Plant RNA Isolation]

Methylation-Specific Digital Karyotyping Protocol - http://www.natureprotocols.com/2006/11/09/methylationspecific_digital_ka.php

Category: Cytogenetics Protocols

Protocol describes a recently developed method — methylation-specific digital karyotyping (MSDK) — that enables comprehensive and unbiased genome-wide DNA methylation analysis. Using a combination of a methylation-sensitive mapping enzyme (for example, AscI) and a fragmenting enzyme (for example, NlaIII), short sequence tags can be obtained and uniquely mapped to genome location. - [Read Methylation-Specific Digital Karyotyping Protocol]

Microarray Analysis of Highly Purified Human Decidual NK Cells Protocol - http://www.natureprotocols.com/2006/08/07/microarray_analysis_of_highly.php

Category: Microarray Protocols: Microarray Analysis Protocols

Description of gene array analysis and normalization - [Read Microarray Analysis of Highly Purified Human Decidual NK Cells Protocol]

Microarray Success - http://www.microarraysuccess.org

Category: DNA Microarray Protocols

The "7 keys to successful microarray analysis" has been designed to guide researchers step-by-step through the design, implementation, analysis, and publication of a microarray experiment. - [Read Microarray Success]

Microarrays a Complete Manual PDF - http://research.nhgri.nih.gov/microarray/Protocols.pdf

Category: DNA Microarray Protocols

Microarray fabrication, coating, slide blocking, printing microarrays, RNA extraction, Sample Labeling for analysis, hybridization, complete protocol. NHGRI - [Read Microarrays a Complete Manual PDF]

Microdissection Overview - http://cgap-mf.nih.gov/Protocols/Microdissection/Overview.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Human tissues are comprised of multiple interacting cell populations in a complex three dimensional arrangement with each cellular phenotype determined by a unique profile of mRNA and protein expression. Before microdissection techniques were developed, the only analysis tools for phenotypic studies were primarily immunohistochemistry and in-situ hybridization. While useful, these tools are limited to single gene analysis and, in general, do not allow qualitative studies. - [Read Microdissection Overview]

Microscopic Analysis of Mitochondrial Transmembrane Potential Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4462

Category: Cell Organelle Protocols: Mitochondria Protocols

Protocol describes a method for the evaluation of mitochondrial function using the fluorochrome CMXRos. CMXRos is sequestered by actively respiring mitochondria, but washed out when the mitochondrial membrane potential is lost. This analysis can be combined with the TUNEL technique or immunocytochemistry. - [Read Microscopic Analysis of Mitochondrial Transmembrane Potential Protocol]

Millipore's ZipTip Protocol - http://www.millipore.com/publications.nsf/docs/tn072

Category: Mass Spectrometry Protocols

Millipore's ZipTip Protocol. Intended for purifying and concentrating femtomoles to picomoles of protein, peptide or oligonucleotide samples prior to analysis, providing better data quality. Millipore. - [Read Millipore's ZipTip Protocol]

Molecular Analysis and Results - DNA Comparative Genomic Hybridization (CGH) - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/DNA/CGH.html

Category: Cytogenetics Protocols: Comparative Genomic Hybridization CGH Protocols

Based on the original protocol describing CGH by Kallioniemi et al. - [Read Molecular Analysis and Results - DNA Comparative Genomic Hybridization (CGH)]

Molecular and Biochemical Analysis of Arabidopsis Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_197-207.pdf

Category: Plant Biology Protocols

The standard protocol for in situ hybridizations in plants still involves fixing fresh tissue, embedding the tissue in wax, sectioning with a microtome and detection of the transcripts of interest using labeled RNA-probes. This protocol concentrates only on nonradioactive methods, as they are easy to perform, very sensitive and even faster than techniques involving radioisotope labels. - [Read Molecular and Biochemical Analysis of Arabidopsis Protocol]

Monosaccharide Analysis by Methanolysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4247

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol describes monosaccharide analysis by methanolysis. The method gives more complex chromatograms than the corresponding alditol acetate method, because up to four methyl glycosides can be produced from each monosaccharide. However, the method is easier to perform. - [Read Monosaccharide Analysis by Methanolysis Protocol]

Monosaccharide Analysis of Phosphorylated Sugars by Methanolysis and Diazomethane Methylation - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4248

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol presents a method that allows rapid determination of the monosaccharide composition of glycans, glycoproteins, and proteoglycans that contain (or are suspected to contain) phosphorylated sugars. (For samples that do not contain phosphorylated sugars see Monosaccharide Analysis by Methanolysis.) - [Read Monosaccharide Analysis of Phosphorylated Sugars by Methanolysis and Diazomethane Methylation]

Monosaccharide Composition Analysis: Alditol Acetates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4246

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol describes how to determine the monosaccharide composition of glycans, glycoproteins, or proteoglycans by hydrolyzing the sample to monosaccharides and converting them to alditols, then performing acetylation to make them volatile compounds and analysis by gas chromatography (GC) or gas chromatography coupled with mass spectrometry (GC-MS). - [Read Monosaccharide Composition Analysis: Alditol Acetates Protocol]

Mouse Castration Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4387

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes castration of male mice, which is used to eliminate testicular hormones and/or to obtain testes for analysis without sacrificing the male. Castration is performed in a similar manner to vasectomy. - [Read Mouse Castration Protocol]

Mouse Nephrectomy Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4386

Category: Mouse Protocols: Mouse Organ Removal Surgery Protocols

Protocol describes surgical removal of a mouse kidney to provide tissue for analysis. - [Read Mouse Nephrectomy Protocol]

Mouse Splenectomy Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4385

Category: Mouse Protocols: Mouse Organ Removal Surgery Protocols

Protocol describes removal of a mouse spleen to provide tissue for analysis. - [Read Mouse Splenectomy Protocol]

Multiparameter FACS Analysis - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C9AB332DE6E4548834730AEA427&objectid=6674F9C194065C413146C81056933A0C

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Provides a brief historical perspective to illustrate the demands for this technology and to lay the foundation for its application; Explains the hurdles that were surmounted to achieve the current level of multiparametric analysis which serves to alert researchers to potential problems they may encounter when they either bring this technology to their own laboratories, or when they use extant instrumentation in another laboratory; Illustrates some of the complexities that arise. - [Read Multiparameter FACS Analysis]

MULTIPLEX SNP ANALYSIS: SCREENING FACTOR V R506Q (LEIDEN) MUTATIONS - http://www.beckmancoulter.com/Literature/BioResearch/A-1928A.pdf

Category: DNA Protocols: SNP Protocols

MULTIPLEX SNP ANALYSIS: SCREENING FACTOR V R506Q (LEIDEN) MUTATIONS. Beckman Coulter. - [Read MULTIPLEX SNP ANALYSIS: SCREENING FACTOR V R506Q (LEIDEN) MUTATIONS]

Multiplexing Cell Viability Assays Protocols - http://www.promega.com/paguide/chap4.htm#title6

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The latest generation of Promega cell-based assays includes luminescent and fluorescent chemistries to measure markers of cell viability, cytotoxicity and apoptosis, as well as to perform reporter analysis. Using these tools researchers can investigate how cells respond to growth factors, cytokines, hormones, mitogens, radiation, effectors, compound libraries and other signaling molecules. The protocols provided are guidelines for multiplexing cell-based assays & are intended as starting points. - [Read Multiplexing Cell Viability Assays Protocols]

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Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

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.

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.

Toluidine Blue Stain

Toluidine Blue staining method and protocol for cell or tissue histology.

Thionin Stain

Thionin staining method and protocol for cell or tissue histology.

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.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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