cultures Protocols

Category: Cytoskeleton Protocols: Actin Myosin Protocols

The same GFP-tagged actin construct used in cell transfection experiments has been used to produce transgenic mice. Transgenic animals allow the imaging of brain tissue in the intact animal, as acutely cut slices or as organotypic slice cultures. They also serve as a source of cells for imaging neurons at high resolution in dispersed low-density cell culture. In contrast to cells transfected in culture, where the level of actin-GFP expression in neurons varies considerably, transgenic mice... - [Read Imaging Actin in Tissue Slices from Transgenic Mouse Brain Protocol]

Category: Western Blot Protocols: General Western Blot Methods Protocols

For immunoblotting experiments, it is often important to compare the total amount of an antigen from many different sources or to learn if a particular source has the antigen under study.  In the approach described here, tissue cultures, bacteria, yeast cells, tissues, and other sources of antigens are disrupted directly in an electrophoresis sample . - [Read Immunoblotting: Preparing Cell Lysates Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

The results of cytotoxicity tests in primary cultures of rat hepatocytes and in MDBK and McCoy cells can be used to predict the in vivo 4-wk maximum tolerated dose in rats and dogs.  A correlation between in vitro cytotoxicity, as measured in this system, and LD50 values in rats and mice has also been established. - [Read In Vitro Prediction of the Maximum Tolerated Dose Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Dilution of Stationary-Phase Cultures Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques

Introduction to Animal Cell Culture. Corning Life Science PDF. Key concepts to Cell Cultures an Introductory Manual with definitions and explanations. - [Read Introduction to Animal Cell Culture - Corning Life Science PDF.]

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol describes isolation of BAC DNA from bacterial cultures using the commercial NucleoBond system. - [Read Isolating BAC DNA from Bacterial Cultures Using the NucleoBond System Protocol]

Category: Mouse Protocols: Mouse Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Isolation and Growth of Primary Cell Cultures from Mouse Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

The procedure for isolation of BAC DNA is scaled-up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BAC DNA. - [Read Isolation of BAC DNA from Large-scale Cultures Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures Protocol]

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for liquid culture of worms. Includes: superbroth; S- basal; worm plates; GROWING THE BACTERIA (WORM FOOD); GROWING THE WORMS; AFTER THE CULTURE HAS GROWN; PREPARING EGGS TO START SYNCHRONIZED LIQUID CULTURES. - [Read Liquid Culture of Worms Protocol]

Category: Tetrahymena Protozoa Protocols

Long-term storage of unfrozen Tetrahymena is convenient when a laboratory is not using the cells frequently; however, cells maintained under these conditions often take a couple of days of incubation in culture medium to recover. - [Read Long-Term Storage of Unfrozen Tetrahymena Cultures in Soybean Medium Protocol]

Category: Tetrahymena Protozoa Protocols

Protocol provides a method for long-term storage of Tetrahymena cultures using maintenance culture medium. - [Read Long-Term Storage Storage of Unfrozen Tetrahymena Cultures in Maintenance Culture Medium Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on Luria-Bertani (LB) agar on or in LB broth containing Ampicillin (30 µg/ml) or Carbenicillin (50 µg/ml broth, 100 µg/ml agar). E. coli strains are usually preserved in stab agar or glycerol for mid-term storage and lyophilized for long-term storage. - [Read Maintenance of Probes in Bacteria Including Escherichia coli Protocol]

Category: Yeast Protocols

Simple method for determining the cell density of yeast cultures. - [Read Measuring Yeast Cell Density by Spectrophotometry Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Membrane permeability of perfused cell cultures, as determined by the efflux of [3H]-2-deoxy-D-glucose-6phosphate, is used as an indicator of the cytotoxic effect of chemicals. - [Read Membrane Permability as a Measure of Cytotoxcity in Perfused Cell Cultures]

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]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Method for Establishment and Maintenance of Leaf and Root Cell Cultures Arabidopsis thaliana. CL ENCINA1, M CONSTANTIN2 and J BOTELLA2. Plant Molecular Biology Reporter 19: 245–248, 2001. - [Read Method for Establishment and Maintenance of Leaf and Root Cell Cultures Arabidopsis thaliana]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Insect cell cultures are now commonly used in insect physiology, developmental biology, pathology, and molecular biology. As the field has advanced from methods development to a standard procedure, so has the diversity of scientists using the technique. This paper describes methods that are effective for maintaining various insect cell lines. The procedures are differentiated between loosely or non-attached cell strains, attached cell strains, and strongly adherent cell strains. - [Read Methods for Maintaining Insect Cell Cultures]

Category: Cytogenetics Protocols

Protocol for mitotic chromosome from cell cultures. Includes: Colchicine Treatment; Monolayer cultures; Cell Suspensions; Cell Harvest; Chromosome Slide Preparation. - [Read Mitotic Chromosome from Cell Cultures Protocol]

Category: Cytogenetics Protocols

Protocol for mitotic chromosome preparations from mammalian suspension cell cultures. - [Read Mitotic Chromosome Preparations from Mammalian Suspension Cell Cultures Protocol]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

One of the most important, but frequently overlooked, cell culture procedures is testing cultures for microbial contamination, especially mycoplasma. It is critical for every cell culture laboratory to only use cell lines that have been carefully screened for mycoplasma. Fortunately, there is a simple fluorochrome DNA staining test that can detect both mycoplasma and virtually any other prokaryote contaminants. - [Read Mycoplasma Detection Using DNA Staining Protocol]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

For both biological and economical reasons, it is important to eliminate mycoplasmas from cell cultures being used for basic research, diagnosis, and biotechnological production. The most commonly used method for elimination, inactivation, or suppression of mycoplasmas in cell cultures is treatment with antibiotics. In general, antibiotic therapies do not result in long-lasting, successful elimination. Also, the cytotoxic properties of antibiotics can cause undesirable side effects on cells. - [Read Mycoplasma Elimination Reagent Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Describes the steps in detail to isolate and expand neural stem cells in the form of neurospheres from tissue dissections of the post-natal mouse brain. Procedures for the long term passage of neurospheres and the cryopreservation of neurospheres are also provided. In addition to the guidelines and tips for generating neurosphere cultures, we describe the method to prepare neurospheres for analysis by light microscopy. - [Read Neural Stem Cell Culture: Neurosphere Generation, Microscopical Analysis and Cryopreservation]

Category: Bacteria Transformation Protocols

Protocol generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). - [Read Preparation and High-efficiency Transformation of Competent E. coli]

Category: E Coli Protocols: E coli Transformation Protocols

Procedure generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). IMPORTANT All steps in this protocol should be carried out aseptically. - [Read Preparation and Transformation of Competent E. coli Protocol]