includes Protocols

Category: Signalling Signal Transduction Protocols

Protocol describing (PI 3-Kinase) activity assay. Includes Preparation of Phosphatidylinositol (PI). - [Read (PI 3-Kinase) Activity Assay]

Category: Immunohistochemistry Protocols: Fixation Protocols

Protocol for 10% formal saine. Includes: 100 litres of 10% Formal Calcium. - [Read 10% Formal Saline Protocol]

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

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol for 3/7 assay. Includes: Detection of Caspase-3 and -7 Activities in Cell-Based Assays; Detection of Caspase-3 or -7 Activity Using Purified Caspases. - [Read 3-7 Assay Protocol]

Category: Immunohistochemistry Protocols: Fixation Protocols

Protocol for 4% paraformaldehyde. Includes: 4% PARAFORMALDEHYDE/PBS; 0.4% PARAFORMALDEHYDE/PBS (FOR POST-DIGESTION FIXATION). - [Read 4% Paraformaldehyde Protocol]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for 96-well confirmation Yeast PCR. Includes: Clonal purification; Generate a master plate (96-well format); Making a frozen backup stock; Confirmation PCR for one Row; ORF Specific Confirmation PCR --> "A-B" primers (upstream junction); Transfer template DNA to multiwell PCR plate; Prepare and dispense master mix for A-B PCR. - [Read 96-Well Confirmation Yeast PCR Protocol]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple samples of organelle DNA from a small amount of tissue. - [Read A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower]

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

Protocol describes typical methods that are used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read A Protocol for AAV Vector Production and Purification]

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

A simplified system for rapid generation of recombinant adenoviruses. Includes: Generation of Recombinants in Bacterial Cells; Viral Production in 293 or 911 Cells; Preparation of High Titer Viral Stocks. - [Read A Simplified System for Rapid Generation of Recombinant Adenoviruses]

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

Describes an experimental cross in mice that can be used to define and map induced germ-line mutations that map to a single chromosome. The cross is a modification and extension of a conventional three-generation recessive mutagenesis screen. Includes: The Mutagenesis Breeding Plan; Consomic Strains; Generating Mutations; Generating and Genotyping G2 Females; Genotyping G3 Progeny; Phenotyping G4 Progeny; etc.. - [Read A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol]

Category: Lipid Protocols: Arachidonic Acid Protocols

Protocol for AA and metabolite quantitation of cell pellets post AA labeling. Includes: Extraction and TLC. - [Read AA & Metabolite Quantitation of Cell Pellets Post-AA Labeling Protocol]

Category: Cloning Protocols: Vector Protocols

Protocol is used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read AAV Vector Production and Purification Protocol]

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for ABI prism 310 cycle sequencing. Includes: Sequencing Reaction; Column Purification; Sample Loading Preparation. - [Read ABI Prism 310 Cycle Sequencing Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

For analysis of metaphase chromosomes, any tissue containing dividing cells can be used: Root tips from young seedlings, from newly grown roots at the edge of plant pots or hydroponic culture are all suitable. Alternatively, flower buds, anthers, carpels or leaf or apical meristems can be used. Includes metaphase arresting reagents. - [Read Accumulation and Fixation of Plant Metaphase Chromosomes Protocol]

Category: Cytoskeleton Protocols: Actin Myosin Protocols

Protocol includes: Binding assays for tropomyosin, villin or caldesmon and Bundling Assays - [Read Actin Binding & Bundling Sedimentation Assays]

Category: Cytoskeleton Protocols: Actin Myosin Protocols

Protocol for actin capture assay. Includes: Actin Capture Assay Reagants. - [Read Actin Capture Assay Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

AFLP was designed as a highly sensitive method for DNA fingerprinting to be used in a variety of fields. We are using this technology to generate DNA based markers for cloning genes involved in phototropic responses in higher plants that have only been identified genetically by mutant phenotype. Protocol includes: Generate polymorphic recombinant F2 (or F3) population; Isolate genomic DNA; Restriction of DNA; Ligation of adapters; Pre-amplification of template DNA; AFLP-PCR; etc. - [Read AFLP For Positional Cloning]

Category: PCR Protocols: AFLP PCR

AFLP Procecure, includes restriction Digestion of Genomic DNA. Michael Havey - Vegetable Genetics Laboratory - [Read AFLP Method]

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

Protocol for agarose gel electrophoresis of DNA. Includes: Making the gel; Loading the gel; Running the gel.
- [Read Agarose Gel Electroporesis of DNA Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the aggregation of ES cells with mouse morula-stage embryos. Includes: Preparation of aggregation walls; ES cell preparation; Embryo preparation and aggregation. - [Read Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol]

Category: Signalling Signal Transduction Protocols

Protocol for Akt/PKB kinase assay. Protocol includes: Akt lysis buffer; Kinase Buffer; 1.5x Akt kinase reaction buffer; Start solution; Lipofectamine PLUS; CDNA synthesis (Roche #1483?88). - [Read Akt/PKB Kinase Assay]

Category: Signalling Signal Transduction Protocols

Technique for Alkaline Phosphatase (APAAP). Techniques includes information on: Cytological Preparations, Fixation, Technique, Results, Substrate Solution and 0.005M Tris/HC1 Saline Buffer pH 7.6. - [Read Alkaline Phosphatase (APAAP) Technique]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for alkaline phosphatase (APAAP) technique. Includes: Fixation; Technique; Substrate Solution. - [Read Alkaline Phosphatase (APAAP) Technique Protocol]

Category: Microarray Protocols: Amino-allyl Microarray Methods

Protocol for amino-allyl labeling (GeneTAC). Includes: RT Reaction; Nucleotide Mix for one reaction; Coupling; Prehybridization of probe. - [Read Amino-allyl Labeling (GeneTAC) Protocol]

Category: Cloning Protocols: Baculovirus Protein Expression Protocols

Protocol for amplification of Baculovirus plaques and preparation of high titer virus stocks. Includes: Primary Amplification; Secondary Amplification. - [Read Amplification of Baculovirus Plaques and Preparation of High Titer Virus Stocks Protocol]