experimental Protocols

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: Protein Protocols: Protein Phosphorylation Protocols

Paper describing methods for monitoring kinase activity, investigating kinase–substrate specificity, examining phosphorylation in planta and the determination of phosphorylation sites in a protein. In addition, strategic considerations for experimental design and variables will be discussed. Scott C. Peck, the Plant Journal. - [Read Analysis of protein phosphorylation: methods and strategies PDF]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Apoptosis: A Laboratory Manual of Experimental Methods. Andrea Cossarizza & Diana Boraschi - [Read Apoptosis: A Laboratory Manual of Experimental Methods]

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for the basic methods of culturing Drosophila. Includes: Stockkeeping; Culture contaminants; Experimental populations. - [Read Basic Methods of Culturing Drosophila Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for calcium flux analysis. Includes: Reagent Preparation; Sample Preparation; Cell Surface Staining; Instrument Set up; Optimizing the Instrument Settings; Recording Data to Apply Compensation; Recording Experimental Data. - [Read Calcium Flux Analysis Protocol for Flow Cytometry]

Category: siRNA and RNAi Protocols

DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE. using T7 to create RNA. Carolyn Worby. Dixon Lab RNAi experiments. - [Read DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE]

Category: Protein Microarray Protocols

Experimental Procedures for Protein Microarrays. MacBeath G & Schreiber SL (2000). Printing proteins as microarrays for high-throughput function determination. Science 289: 1760-1763. - [Read Experimental Procedures for Protein Microarrays]

Category: Real Time PCR

A detailed experimental procedure for real-time RT-PCR using SYBR Green I as mentioned in Wang et al 2003. - [Read Protocol for Real-Time RT-PCR Using SYBR Green]

Category: siRNA and RNAi Protocols

RNA interference protocol. Preparation of Template DNA, Preparation of the dsRNA, RNAi in Drosophila Schneider cells. Matthias Wilm. - [Read RNA interference protocol]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments.  This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos.  The tissue is cut in slices using a vibratome or tissue slicer.  The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. - [Read Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Category: Avian Bird Protocols

Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. The tissue is cut in slices using a vibratome or tissue slicer. The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. - [Read Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Simple Experimental Method for Storage of Cell Lines. Hancock Lab. - [Read Storage of Cell Lines - Hancock]

Category: Transfection Protocols

Transfection of IMEC cells using SuperFect Transfection Reagent protocol. Protocol includes experimental design considerations. - [Read Transfection of IMEC Cells Using SuperFect Transfection Reagent Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase assay]

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase Assay Protocol]

Category: Plant Biology Protocols

Alfalfa is an outcrossing species, cultivars consist of populations rather than individual homozygous or inbred lines. After only two cycles of self-pollination, severe inbreeding depression eliminates selfed individuals from populations. To obtain sufficient plant material of relative genetically uniformity (these plants will still necessarily be heterozygous) for experimental purposes, it may be necessary to propagate a single individual or clone. - [Read Vegetative Propagation of Alfalfa by Stem Cuttings]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments.  This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos.  This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation). - [Read Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Category: Avian Bird Protocols

Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation). - [Read Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol]