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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]

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]

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

Culture of Human Embryonic Stem Cells (hESC) - http://stemcells.nih.gov/research/NIHresearch/scunit/culture.asp

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Culture of Human Embryonic Stem Cells (hESC). one simple protocol outlined. NIH Stem Cell Unit - [Read Culture of Human Embryonic Stem Cells (hESC)]

Fragmentation of DNA by Nebulization Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4539

Category: Cloning Protocols

Protocol describes a method for DNA fragmentation by nebulization, in which the fine mist created by forcing a DNA solution through a small hole in the nebulizer unit is collected. The size of the fragments obtained by nebulization is determined chiefly by the speed at which the DNA solution passes through the hole, altering the pressure of the gas blowing through the nebulizer, the viscosity of the solution, and the temperature. - [Read Fragmentation of DNA by Nebulization Protocol]

Measurement of Apoptosis and Other Forms of Cell Death - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6636C9D7EA88169E130B219EADFA39&objectid=6674C718AE37D97A78A88E365485C689

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

Measurement of Apoptosis and Other Forms of Cell Death. Jagan Muppidi, Melissa Porter, and Richard M. Siegel. As programmed cell death (PCD) or apoptosis has emerged as an important regulator of development and homeostasis in multicellular organisms, methods to quantify apoptosis and to distinguish it from necrosis have been developed. This unit presents a set of assays for these purposes, many of which are technically very simple and ideally suited to the study of hematopoietic cells. - [Read Measurement of Apoptosis and Other Forms of Cell Death]

Media and Culture of Yeast Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E5192D47CE9B0776DB7D136C34ED148A&objectid=6673ABA6C2DB2E4E7564EDFA26589256

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

The yeast, Saccharyomyces cerevisiae, has become an important organism in molecular, biochemical, and genetic analysis. The organism has specific requirements for growth under a variety of conditions. The media, both liquid and solid, simple, define, and complex are describe in this unit. Also included are methods for handling, storing, and shipping stock of yeast. - [Read Media and Culture of Yeast Protocol]

Sterilization and Filtration Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662B9D0482A943EC97AC4807854493&objectid=6673A220D50910E56F81295AE515690F

Category: Cell Culture Protocols: Sterile Technique Protocols

This unit on sterility in the tissue culture environment describes methods for sterilization of liquid and dry goods used for tissue culture and filtration of liquids to prevent contamination of cultures. - [Read Sterilization and Filtration Protocol]

Subcloning Protocol ES Cells - http://stemcells.nih.gov/research/NIHresearch/scunit/subcloning.asp

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Subcloning Protocol ES Cells. Used an adapted form of the feeder-free protocols detailed in Xu et al. (Nature Biotechnology 19:971â€“974, 2001). NIH Stem Cell Unit - [Read Subcloning Protocol ES Cells]

T Cell Enrichment by Cytotoxic Elimination of B Cells and Accessory Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66364CDDD21842657A5068627A2F0C&objectid=667495E7DA47001CA835F427F96D1C3B

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Describes T cell enrichment using cytotoxic antibodies, and also describes the depletion of T cells and their subpopulations using the same approach. In the latter unit, T cell surface markers (Thy-1, CD4, and CD8) are targeted by the cytotoxic antibodies. - [Read T Cell Enrichment by Cytotoxic Elimination of B Cells and Accessory Cells Protocol]

Articles (1-5 of 5)

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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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