growth Protocols

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Aggregations - Production of Chimerae

Aggregation: Production of Chimerae. Detailed ES cell growth and preparation information for aggregations. Edward W. Scott Univ. Florida Shands Cancer Center - [Read Aggregation: Production of Chimerae]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: Plant Biology Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: DNA Protocols: Cosmid Library Protocols

Amplification of cosmid libraries may result in distorted representation of cloned genomic sequences and should be avoided wherever possible. If amplification should become necessary, it is best to expand the library by growth in liquid medium. - [Read Amplification and Storage of a Cosmid Library: Amplification in Liquid Culture Protocol]

Category: Neuroscience Protocols

After section of the corticospinal (CS) tract in adult rats, neutralizing Nogo-A with monoclonal antibodies leads to enhancement of axonal re-growth and compensatory sprouting, accompanied by increased motor recovery.  The neutralization of Nogo-A represents a promising approach for therapy after lesion if its enhancement of functional recovery can be transposed to primates. - [Read Anti-Nogo-A Treatment Protocol]

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: Reporter Gene Assays: B-Galactosidase Assay Protocols

There are two basic methods for the in vitro assay of B-galactosidase activity from yeast. They differ mainly in the method of preparing the material for assay. Both methods are described with accompanying protocols. Method I: Assay of Crude Extracts includes: Yeast Cell Growth; Yeast Cell Harvest; B-gal assays; Bradford Assays. Method II: Permeabilized cell assay. - [Read Assay of β-Galactosidase in Yeast Protocol]

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

Protocol for assay of β-Galactosidase in Yeast. Includes: Assay of Crude Extracts; Yeast Cell Growth; Yeast Cell Harvest; β-gal assays; Bradford Assays; Permeabilized cell assay. - [Read Assay of β-Galactosidase in Yeast Protocol]

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

Protocol for balanced salts used for growth media. - [Read Balanced Salts for Growth Media Protocol]

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

Protocol provides methods for cryofreezing and subsequent thawing of mammalian cells. Pre-confluent cells are trypsinized, pelleted, resuspended in freezing medium, and gradually frozen. When needed, frozen cells are thawed quickly under running tap water and transferred to growth medium. - [Read Cryopreservation of Mammalian Culture Cells: Preparation and Recovery of Samples Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol utilizes the Y190 yeast strain, that characteristically turns from white to red during growth due to ademutation. Baits were subcloned into the pAS1cyh2 vector, and Preys subcloned into pACT2. - [Read Directed Yeast Two Hybrid Screening for MEKK1 Interaction Partners Protocol]

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila growth conditions. Includes: MAINTENANCE; Semi-adherent cell lines; Adherent cell lines; SL2, S2C, S2*; DL2, DL1, S2R+, Kc167, Clone 8, S3; BG2. - [Read Drosophila Growth Conditions Protocol]

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

Protocol for eagle growth media (DME). - [Read Eagle’s Growth Media (DME) Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Reference: Michael P. Matise, Wotjek Auerbach and Alexandra L. Joyner (2000). Gene targeting: a practical approach. Protocol excerpted from Chapter 3, Production of targeted embryonic stem cell clones. Alexandra L. Joyner (ed.), 2nd edition, Oxford Unive - [Read Embryonic Stem Cell growth media Requirements - Taconic Transgenics]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: Embryotoxic Potential Testing of Chemicals

EMBRYONIC STEM CELL TEST (EST). The embryotoxic potential of chemicals is determined by the evaluation of the inhibition of differentiation of embryonic stem cells (ES) and the inhibition of growth of ES and 3T3 cells. Scientific Information Service - [Read EMBRYONIC STEM CELL TEST (EST)]

Category: Cell Biology and Cell Culture

Eukaryote Growth Dynamics. Cell Biology Laboratory Manual. Heidcamp. - [Read Eukaryote Growth Dynamics]

Category: Bacterial Cell Culture Protocols

The growth conditions of microbial cell cultures and the time of sample collection should be optimized and standardized when growing cells for protein extraction. Because cells may excrete proteases and other extracellular enzymes, and compounds in the medium may interfere with extraction, wash cultures with an isotonic buffer, such as PBS or sucrose before solubilization. - [Read Extraction and Solubilization of Total Protein from Microorganisms 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: Fungi Protocols: Aspergillus Protocols

Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc - [Read Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for fungal DNA isolation. The key elements in this prep are (1) the use of young lyophilized mycelial mats....young mats (4 days growth for C. carbonum)...yield less contaminating carbohydrates and other misc. junk (2) lots of proteinase K in the extraction buffer to kill Dnases (final =0.3mg/ml). - [Read Fungal DNA Isolation Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: S Phase Protocols

This protocol provides a method for synchronizing cells at the G1/S border using a double treatment of thymidine, which, in excess, is an inhibitor of DNA synthesis. Cells are treated once with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. - [Read G1/S Phase Synchronization using Double Thymidine Synchronization Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol provides a method for synchronizing cells at the G1/S border using a double treatment of thymidine, which, in excess, is an inhibitor of DNA synthesis. Cells are treated once with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. - [Read G1/S Phase Synchronization using Double Thymidine Synchronization Protocol]