arabidopsis Protocols

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Experiments included for analysis of plant epidermis: Steroid induction of gene expression in Arabidopsis; Replica Molds and Casts of the plant epidermis. - [Read Analysis of the Plant Epidermis Experiments]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Procedure to isolate DNA from Arabidopsis Thaliana. - [Read Arabidopsis Thaliana DNA Isolation Procedure]

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Protocol for Arabidopsis thaliana DNA isolation procedure. - [Read Arabidopsis thaliana DNA Isolation Protocol]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Arabidopsis Tissue Culture Stock Solutions and Media. Lazo Lab. MS MEDIUM FOR ARABIDOPSIS. B5 MEDIUM FOR ARABIDOPSIS. 10X Micronutrients. - [Read Arabidopsis Tissue Culture Stock Solutions and Media]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Electrotransformation of Agrobacterium with a plasmid that has been replicating in E. coli. Growth of Arabidopsis thaliana. Arabidopsis dunking. Seed Harvesting. Plant tissue culture. Very detailed protocol. Stockinger lab. PDF - [Read Arabidopsis transformation with Agrobacterium PDF]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Arabidopsis transformation and selection on kanamycin agar, Preparation of plants for transformation, Agrobacterium, Dipping, Post-dip care, Agar plates, Tips. Adaptation of the short protocol described bySteve Clough and Andrew Bent, University of Illinois at Urbana-Champaign. - [Read Arabidopsis transformation with pictures]

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]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Method is for preparing chromosomes from single flower buds of A. thaliana. It does not kill the plants allowing the determination of their chromosome number throughout development. Includes: Preparations of Arabidopsis; Preparation of Chromosome; Staining Chromosomes. - [Read Chromosome Spreads from Flower Buds of Arabidopsis thaliana Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact non-ovule materials of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse root, seedling even flower development without sectioning. This protocol could also be used for clearing GUS stained material, after chlorophyll is removed by 70% ethanol. - [Read Clearing Arabidopsis Non-Ovule Materials With the HCG Solution Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact seeds of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse embryo and endosperm development without sectioning. - [Read Clearing Arabidopsis Ovule Materials with the Hoyer's Solution Protocol]

Category: Plant Biology Protocols

In most natural habitats, Arabidopsis is a winter annual: Its seeds germinate in the fall, the young plants survive the winter, floral meristems emerge in the spring, and only the seeds survive the summer months. Most common laboratory varieties of Arabidopsis flower within 4 weeks of germination, and seeds can be collected after an additional 4-6 weeks. - [Read Cultivation of Arabidopsis Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

EMS is used at concentrations that induce multiple point mutations in each plant, such that mutant alleles of a specific locus are found at a rate of ~1 in 2000-5000 M2 plants. This high rate of mutagenesis makes possible the screening of relatively few plants to find those with the phenotype of interest, a particular advantage if the screen is laborious or if only a small number of genes mutate to the required phenotype. - [Read EMS Mutagenesis of Arabidopsis Seed Protocol]

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]

Category: Plant Biology Protocols

Protocol for the fixation and seperation of Arabidopsis leaf cells. Includes: Sample Preparation; Slide Preparation. - [Read Fixation and Separation of Arabidopsis Leaf Cells Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Forward genetics is used to identify genes that are involved in particular biological processes. For example, genes required for disease resistance can be found by identifying mutants with reduced or increased disease resistance, genes that control flower development can be identified by searching for mutants with altered flower morphology, and genes encoding enzymes for tryptophan biosynthesis can be identified by searching for mutants that require exogenous tryptophan for growth. - [Read Forward Genetics in Arabidopsis: Finding Mutations that Cause Particular Phenotypes Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for genetic mapping of Arabidopsis centromeres using PCR-based markers. Includes: DNA markers facilitate genome analysis; Centromere mapping using tetrad analysis; Preparation of DNA from frozen plant tissue. - [Read Genetic Mapping of Arabidopsis Centromeres Using PCR-Based Markers Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol describes the use of glufosinate ammonium to select transformed Arabidopsis plants. The major advantage of glufosinate ammonium selection is that it can be performed on plants growing in soil and does not require the use of sterile techniques. - [Read Glufosinate Ammonium Selection of Transformed Arabidopsis Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol on how to stain Arabidopsis meiotic chromosomes from pollen meiocytes. - [Read How to Stain Arabidopsis Meiotic Chromosomes From Pollen Meiocytes Protocol]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for a rapid and robust method of identifying transformed Arabidopsis thaliana seedlings following floral dip transformation. - [Read Identifying Transformed Arabidopsis thaliana Seedlings Following Floral Dip Transformation Protocol]

Category: Microscopy Protocols: Electron Microscopy Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope]

Category: Plant Biology Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope Protocol]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

The plant transformation procedures described involve floral dip, vacuum infiltration, and spraying. They yield transformants at frequencies ranging up to several percent, with the most common frequency being 0.1%-1%. - [Read In Planta Transformation of Arabidopsis Protocols]

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Includes: Isolation of Arabidopsis LEU2 cDNAs by complementation of the yeast leu2 mutation; Recovery of plasmid DNA from yeast cells; Analysis of Arabidopsis cDNAs that complement a yeast leu2 mutation; Preparation of library DNA from the Lacroute cDNA library; Preparation of Yeast Media. - [Read Isolation of Arabidopsis cDNAs by Complementation in Yeast]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]