plant Protocols

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

A. thaliana has a very small haploid genome and this makes obtaining DNA somewhat difficult. The most notable problem is that DNA is usually contaminated with polysaccharide which inhibit restriction enzymes as well as other DNA modifying enzymes. This problem is most easily solved by using young plants which have not accumulated as much polysaccharide as older plants. The best results are obtained with plants that are two to three weeks post germinated. - [Read Plant DNA Extraction Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Plant Cell Culture Growth Media Protocols

Protocol for plant growth media. - [Read Plant Growth Media Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Plant Cell Culture Growth Media Protocols

Protocol for plant pathology media. - [Read Plant Pathology Media Protocol]

Category: Protein Protocols: Protein Extraction From Tissue

Plant Protein Isolation - Rapid isolation of protein for SDS-PAGE analysis. (Essentially the same protocol as that described for GUS Assays). Stockinger Lab - [Read Plant Protein Isolation]

Category: Plant Biology Protocols: Plant Tissue Culture

Integration of plant tissue culture into plant. transformation protocols. Various methods of plant regeneration are available to the plant biotechnol- ... - [Read Plant tissue culture]

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Plant Cell Culture Growth Media Protocols

Protocol for plant tissue culture media. Includes: Preparation from Packaged Powder; Preparation from Basal Salt Solutions. - [Read Plant Tissue Culture Media Protocol]

Category: Cell Biology and Cell Culture

Plant Tissue Culture Protocols. Sigma - [Read Plant Tissue Culture Protocols. Sigma]

Category: Plant Biology Protocols: Plant Tissue Culture

Plant tissue culture techniques. Pages 151-174, in Tested studies for laboratory ... engineering protocols employ callus initiation procedures after DNA has ... Mineo Zoology Univ. Toronto - [Read Plant Tissue Culture Techniques]

Category: E Coli Protocols: E coli Genetics Protocols

Protocol for precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. Describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in E coli Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

Describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. Includes: Key steps in the EL250 RED-HR locus rescue and engineering procedure; Primer design and plasmid constructs; AtSTM gap-repair construct; Targeting construct backbone; Preparation of electrocompetent EL250 cells; Transformation of BAC F24o1 and induction of recombinogenic function in EL250; AtSTM locus rescue from BAC F24o1 by gap-repair HR. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Tissues are homogenized in guanidinium isothiocyanate lysis buffer and then subjected to cesium chloride ultracentrifugation. The resulting RNA is then extracted with phenol and chloroform to remove protein and lipid contaminants. - [Read Preparation of RNA from Plant Tissue Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifug]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol is based on the standard Trizol protocol for the purification of RNA from animal cells using Trizol (Purification of RNA from Animal Cells using Trizol). In this version, adapted for use with plant tissues, a high-salt isopropanol precipitation step has been added to precipitate RNA selectively, while maintaining polysaccharides and proteoglycans in solution. - [Read Preparation of RNA from Plant Tissue Using Trizol]

Category: Protein Protocols: Protein Extraction Protocols

Protocol for Protein Extraction Using Proteomics. Extraction of proteins from plant cells that are rich in compounds that interfere with the 2-Dimensional electrophoretic separation methods such as salts, organic acids, phenolics, pigments, terpenes, among others. A common protocol used in our lab for extraction proteins from plant tissues consists in the homogenization of mortar-grounded material in liquid nitrogen with an extraction buffer. - [Read Protocol for Protein Extraction Using Proteomics]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

This protocol is not phenol-based, but does require the addition of chloroform. The Concert Plant Reagent is intended for the isolation of RNA from a wide variety of plant tissues including blue spruce needles, potato tuber etc. - [Read Purification of RNA from Plant Tissue Using the Concert Plant Reagent]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

The activity of ß-glucuronidase (GUS) can be accurately determined in intact plant tissue using 4-methylumbelliferyl ß-D-glucuronide (4-MUG) as a substrate. Upon hydrolysis by GUS, the fluorochrome 4-methyl umbelliferone (4-MU) is produced. This method is based on the permeability of both 4-MUG and 4-MU through plant tissue. It consists of incubation of the tissue with the reagent and quantification of the fluorescence emitted by 4-MU in the solution. GUS activity in each sample can be... - [Read Quantitative GUS Activity Assay in Intact Plant Tissue Protocol]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

Using excitation at 365 nm and measuring emission at 455 nm, the amount of 4-MU produced can be quantified. Under these conditions, background fluorescence from the substrate is negligible, especially if the appropriate filter is selected. - [Read Quantitative GUS Activity Assay of Plant Extracts]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for a simplified Arabidopsis transformation. Found that the MS salts, hormone, etc. make no difference, that OD of bacteria doesn't make much of a difference, that vacuum doesn't even make much of a difference as long as you have a decent amount of surfactant present. Plant health is still a major factor - healthy fecund plants make a big difference! With this method you should be able to achieve transformation rates above 1%. - [Read Simple Arabidopsis Transformation Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

Protocol for the preparation of plant chromosomes using squash. - [Read Squash Preparation of Plant Chromosomes Protocol]

Category: Cytogenetics Protocols

Protocol for squash preparation of plant chromosomes. - [Read Squash Preparation of Plant Chromosomes Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Plant Cell Culture Growth Media Protocols

Protocol for sterilization of plant culture media. Includes: Minimum Autoclaving Time for Plant Tissue Culture Medium. - [Read Sterilization of Plant Culture Media Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]

Category: Protein Protocols: Protein Trafficking

Background and methods to study plant transport. Includes new methods to study protein trafficking in plant cells, includes: Identification of protein sorting pathways in non purified samples; Localization of organelle proteins by isotype tagging/isotype-coded affinity tag; Coupling of chemical genomics and proteomics; Top down mass spectrometry; Compartment-specific markers to aid in the purification of organelles. - [Read Understanding Protein Trafficking in Plant Cells Through Proteomics]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for vacuum-infiltration transformation of Arabidopsis thaliana. Includes: Plant Growth; Vacuum Infiltration; Selection of Putative Transformants; Kanamycin selection. - [Read Vacuum-Infiltration Transformation of Arabidopsis thaliana Protocol]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

A. tumefaciens is a soil-dwelling bacterium that transforms normal plant cells into tumor-forming cells by inserting a piece of bacterial DNA (the transfer, or "T," DNA) into the plant cell genome. The Ti plasmid also carries many of the transfer functions for mobilizing the T-DNA. This article provides a brief discussion of the principles of T-DNA transformation, including consideration of T-DNA vectors and their hosts. - [Read Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol]