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The Isolation and Culture of Rat Hepatic Cells Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/7.htm

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

This method enables the isolation of different populations of liver cells and describes their subsequent culture. - [Read The Isolation and Culture of Rat Hepatic Cells Protocol]

Tissue Culture Methods - http://www.research.umbc.edu/~jwolf/method5.htm#tc%20methods

Category: Cell Biology and Cell Culture: Cell Culture Techniques

Tissue Culture Methods. Julie B. Wolf, Univ. Maryland. Protocols for Counting Cells, Preservation and storage of cell lines, maintenance of cell lines, media and growth, and safety guidelines for working with cells. - [Read Tissue Culture Methods]

Topo Cloning Procedure - http://preuss.bsd.uchicago.edu/protocols/topo.html

Category: DNA Protocols: TOPO Cloning

Topo Cloning procedure for cloning of small amounts DNA fragments by PCR. Cloning the PCR fragment into the TOPO vector, transforming E. Coli cells, and using blue/white selection to determine transformants. Culture inoculation, Qiagen Plasmid Prep protocol for plasmid isolation of the plasmid containing the cloned copies. Preuss Lab, Univ. Chicago. - [Read Topo Cloning Procedure]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. The calcium phosphate method can be used to transfect neurons that have already been growing on coverslips in vitro. Transfected cells suitable for imaging can be obtained in cultures up to 15 days in vitro. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols

Protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol]

Transient Transfection and Luciferase assay - http://www.natureprotocols.com/2006/10/27/transient_transfection_and_luc.php

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]

Transient Transfection and Luciferase Assay Protocol - http://www.natureprotocols.com/2006/10/27/transient_transfection_and_luc.php

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transposon Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for transposon mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon Mediated Mutagenesis Protocol]

Transposon-Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Bacteria Genetics Protocols

Protocol for transposon-mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon-Mediated Mutagenesis Protocol]

Trypsinization of Cells Grown in Monolayer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4349

Category: Cell Culture Protocols: Trypsinization Protocols

Protocol describes the trypsinization of cells in monolayer culture to facilitate subculture or harvesting. To avoid cross-contamination of cells, it is important for each cell line to be subcultured independently. No more than one cell line should be in the tissue culture hood at any one time. - [Read Trypsinization of Cells Grown in Monolayer Protocol]

Understanding and Managing Cell Culture Contamination Protocol - http://www.corning.com/Lifesciences/technical_information/techDocs/cccontamination.pdf

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

No cell culture problem is as universal as that of culture loss due to contamination. All cell culture laboratories and cell culture workers have experienced it. Culture contaminants may be biological or chemical, seen or unseen, destructive or seemingly benign, but in all cases they adversely affect both the use of your cell cultures and the quality of your research. Contamination problems can be divided into three classes: Minor annoyances, Serious problems, Major catastrophes. - [Read Understanding and Managing Cell Culture Contamination Protocol]

V79 Cytotoxicity Test for Membrane Damage - http://embryo.ib.amwaw.edu.pl/invittox/prot/39.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The cytotoxic effect of test chemicals in V79 cell culture can be determined by assessing damage to the plasma membrane as determined by a nucleic acid leakage assay. - [Read V79 Cytotoxicity Test for Membrane Damage]

Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System - http://www.natureprotocols.com/2006/09/15/western_blot_analysis_of_subce.php

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol describes nuclear and cytoplasmic fractionation of tissue culture cells and a method for Western blot detection of proteins using the Odyssey Infrared Imaging System. This protocol was used to detect expression of the "small" Tap protein in 293T, HeLa and COS cells. The Odyssey system has several advantages over the more widely used chemiluminescent detection methods. - [Read Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System]

Yeast DNA Isolation: Midiprep Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4147

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol for Yeast DNA isolation. Protocol describes the preparation of DNA from a 40-ml liquid culture. - [Read Yeast DNA Isolation: Midiprep Protocol]

Yeast DNA Isolation: Miniprep Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4148

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol describes the preparation of DNA from a 5-ml liquid culture of yeast. - [Read Yeast DNA Isolation: Miniprep Protocol]

Yeast RNA Isolation: Large-Scale Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4154

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast RNA Isolation Protocols

Protocol describes large-scale yeast RNA isolation. It is designed to yield more than 10 mg of total nucleic acid and 300-400 µg of poly(A)-selected mRNA from each 500-ml culture. - [Read Yeast RNA Isolation: Large-Scale Protocol]

Yeast Two-Hybrid Screen with Library and Bait - http://faculty.virginia.edu/mirmira/resources_files/Protocols/YEAST%20TWO.htm

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

yeast transformed with bait vector. yeast culture. selection plates. Mirmira, Univ. Virginia. - [Read Yeast Two-Hybrid Screen with Library and Bait]

[3H]Thymidine Labeling Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/T3.html

Category: Cell Culture Protocols

Protocol on [3H]Thymidine labeling. Protocol includes: Explant Culture Medium; 100X (cold) Thymidine; Dektol Developer; Fixer - [Read [3H]Thymidine Labeling Protocol]

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Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

Histone H1 Kinase Activity Assay

Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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