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A Safe Method of Extracting DNA from Coccidioides immitis Protocol - http://www.fgsc.net/fgn42/burt.html

Category: Fungi Protocols: Fungi Genetics Protocols

Protocol describes a safe and convenient method of extracting DNA from Coccidioides immitis fungi in which the culture is killed by steaming, allowing removal from the containment facilities, as soon as possible. The method was first developed with the non-pathogen Neurospora crassa, has worked well for both C. immitis and H. capsulatum, and should be useful for extracting DNA from any pathogenic fungus. - [Read A Safe Method of Extracting DNA from Coccidioides immitis Protocol]

Accumulation and Fixation of Plant Metaphase Chromosomes Protocol - http://www.le.ac.uk/bl/phh4/proaccum.htm

Category: Plant Biology Protocols: Plant Genetics Protocols

For analysis of metaphase chromosomes, any tissue containing dividing cells can be used: Root tips from young seedlings, from newly grown roots at the edge of plant pots or hydroponic culture are all suitable. Alternatively, flower buds, anthers, carpels or leaf or apical meristems can be used. Includes metaphase arresting reagents. - [Read Accumulation and Fixation of Plant Metaphase Chromosomes Protocol]

Adaptation of High Five™ Cells to HyQ© SFX Insect Medium - http://www.hyclone.com/pdf/procedure_insect_highfive.pdf

Category: Cell Culture Protocols: Insect Cell Culture Protocols

This protocol is designed to adapt High Five™ (BTI-Tn-4) cells to serum-free suspension culture in HyQ SFX-Insect in 5 passages. The entire adaptation process takes approximately two weeks and the resulting adapted cells have doubling times of 16 and 20 hours and minimal clumping in suspension culture. - [Read Adaptation of High Five™ Cells to HyQ© SFX Insect Medium]

Agrobacterium-Mediated Transformation in Rice Protocol - http://www.natureprotocols.com/2007/01/18/a_protocol_for_agrobacteriumme.php

Category: Cytogenetics Protocols

Protocol for Agrobacterium-mediated transformation in rice. Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2–3 months with a transformation frequency of 30–50%, both in easily cultured varieties and recalcitrant elite japonica rice. - [Read Agrobacterium-Mediated Transformation in Rice Protocol]

Amplification and Storage of a Cosmid Library: Amplification in Liquid Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4001

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]

An Image-Based Assay of Endothelial Cell Tube Formation as a Model of Angiogenesis - http://www.bdbiosciences.com/pdfs/whitePapers/06-A790030-3A1.pdf

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Traditional animal models to quantify the degree of blood vessel formation are being replaced by cell culture assays that are easier to set up, statistically reliable and can be automated in a drug screening laboratory. These assays rely on the endothelial cells’ ability to form distinct blood-vessel-like tubules in an extracellular matrix where they can subsequently be visualized by fluorescence microscopy. - [Read An Image-Based Assay of Endothelial Cell Tube Formation as a Model of Angiogenesis]

An Integrative Procedure For Apoptosis Identification And Measurement - http://www.natureprotocols.com/2006/08/03/an_integrative_procedure_for_a.php

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

An Integrative Procedure for Apoptosis Identification and Measurement. Yingyu Cui Lab/Group: National Key Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences. I have uploaded an integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read An Integrative Procedure For Apoptosis Identification And Measurement]

Apoptosis Identification and Measurement Protocol - http://www.natureprotocols.com/2006/08/03/an_integrative_procedure_for_a.php

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

An integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read Apoptosis Identification and Measurement Protocol]

Application of Direct Current Electric Fields to Cells and Tissues in vitro - http://www.natureprotocols.com/2006/12/12/application_of_direct_current.php

Category: Cell Biology and Cell Culture

Protocol applies EFs to cells in vitro but has been modified and to use electrotactic chambers to accommodate cells growing in planar culture or in three-dimensional (3D) gels, en bloc tissue cultures in 3D and possible small embryos, such as that from frog and zebra fish. The EF is applied to the cells or tissues cultured in a customer designed electrotactic chamber via agar salt bridges, Steinbergâ€™s solution and Ag/AgCl electrodes. - [Read Application of Direct Current Electric Fields to Cells and Tissues in vitro]

Arabidopsis Tissue Culture Stock Solutions and Media - http://wheat.pw.usda.gov/~lazo/methods/lazo/tissue.html

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]

Arabidopsis transformation with Agrobacterium PDF - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/ArabidopsisTransformation.pdf

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]

Aseptic Technique for Cell Culture Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662B7EFFEB261151E9039B66FD5981&objectid=66739FA0D0CABBF1C466C9F466ABDF9A

Category: Cell Culture Protocols: Sterile Technique Protocols

This unit describes some of the ways that a laboratory can deal with the constant threat of microbial contamination in cell cultures. A protocol on aseptic technique is described first. This catch-all term universally appears in any set of instructions pertaining to procedures in which noncontaminating conditions must be maintained. - [Read Aseptic Technique for Cell Culture Protocol]

Assay of Cytokines in Tissue Culture Supernatants - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000223.pdf?pid=PP00000223

Category: Signalling Signal Transduction Protocols

Assay of cytokines in tissue culture supernatants describes a liquid suspension array for quantification of cytokines in tissue culture supernatants or serum. With this assay, it is possible to profile the level of multiple cytokines in a single well. The principle of this cytokine assay is similar to a capture sandwich immunoassay. Includes: Preparation for the Assay, Cytokine Assay, Reagents and Materials. - [Read Assay of Cytokines in Tissue Culture Supernatants]

Assay of Cytokines in Tissue Culture Supernatants Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000223.pdf?pid=PP00000223

Category: Immunology: Immunoassays

The Bio-Plex cytokine assay employs a liquid suspension array for quantification of cytokines in tissue culture supernatants or serum. Using this 96-well microtiter plateformatted assay, it is possible to profile the level of multiple cytokines in a single well. - [Read Assay of Cytokines in Tissue Culture Supernatants Protocol]

Assessing and Controlling Microbial Contamination in Cell Cultures Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662BAD9F445C2107BC142E6B0F41B0&objectid=6673A7AEBD864E91AC365EE3D108411A

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

This unit describes how to detect and recognize bacterial, fungal, or mycoplasma contamination and how to verify the presence of mycoplasma. Strategies are described for dealing with culture contamination, if necessary. - [Read Assessing and Controlling Microbial Contamination in Cell Cultures Protocol]

Bacterial Glycerol Stocks Protocol - http://www.unc.edu/depts/van_dyke/protocols/bacteria/bacterial_glycerol_stocks.pdf

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

To 2mls of mid-log culture or 1ml of freshly saturated culture add 1ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in liquid nitrogen or on dry ice. Store at -20°C and - 70°C. - [Read Bacterial Glycerol Stocks Protocol]

Basic Cell Culture Protocols - http://www.hyclone.com/library/basicprotocols.htm

Category: Cell Biology and Cell Culture

Basic Cell Culture Protocols. Passaging, Counting, Viability and more. Hyclone - [Read Basic Cell Culture Protocols]

Basic Methods of Culturing Drosophila Protocol - http://fly.bio.indiana.edu/Fly_Work/culturing.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for the basic methods of culturing Drosophila. Includes: Stockkeeping; Culture contaminants; Experimental populations. - [Read Basic Methods of Culturing Drosophila Protocol]

Basic Techniques for Mammalian Cell Tissue Culture Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662B5FE053DEE8CB4AC3EB6F729A11&objectid=667396C6F0D96C40EB68A3D8DB8B201A

Category: Cell Culture Protocols: Sterile Technique Protocols

Cultured mammalian cells are used extensively in cell biology studies; it requires a number of special skills in order to be able to preserve the structure, function, behavior and biology of the cells. This unit describes the basic skills required to maintain and preserve cell cultures: aseptic technique, medium characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. - [Read Basic Techniques for Mammalian Cell Tissue Culture Protocol]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

BHK-21. Basic Cell Culture Protocols - http://www.sci.sdsu.edu/classes/chemistry/chem467l/mardahl/basiccell.html

Category: Cell Biology and Cell Culture

Basic Cell Culture Protocols. BHK-21. - [Read BHK-21. Basic Cell Culture Protocols]

Calcium Phosphate Transfection of Neurons in Primary Culture - http://gweb1.ucsf.edu/labs/finkbeiner/Protocol/protocol_list/Ca_P_TX_R.shtml

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Detailed Neuronal cell transfection protocol and methods. Steven Finkbeiner. UCSF. - [Read Calcium Phosphate Transfection of Neurons in Primary Culture]

Calibration of Micropipette Tips Protocols - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4655

Category: Microinjection Protocols: Microinjection Equipment

This protocol describes an easy method for calibrating micropipette tips that have been pulled in the laboratory. It is essential to estimate the internal diameter of the pulled micropipette tip when adjusting parameters for a new puller or new type of glass tubing. A tip diameter of ~0.3 µm is optimal for the microinjection of mammalian cells in culture (e.g., CHO, PtK1, and COS-7). A 10% increase in diameter increases the delivery rate by more than 30% and can cause cell damage. - [Read Calibration of Micropipette Tips Protocols]

Cell Culture Guidelines - http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_Facility/CellCultureGuidelines.htm

Category: Cell Biology and Cell Culture: Troubleshooting Cell Culture Problems

Immunology Applications Core Facility, Univ. Chicago - [Read Cell Culture Guidelines]

Cell Culture Phototoxicity Test Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/44.htm

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

Human A431 cells and mouse 3T3 cells are exposed in culture to UV light both in the presence and absence of test compound. Phototoxicity is expressed as a decrease in cell viability as determined by the MTT assay. - [Read Cell Culture Phototoxicity Test Protocol]

Articles (1-7 of 7)

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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