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A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./genedel.html~main

Category: Fungi Protocols: Aspergillus Protocols

The technique makes use of an Escherichia coli strain expressing the redΑßΓ operon under the control of an inducible promoter. This enables the strain to carry out homologous recombination with only 50-60 bp of homologous sequence. The procedure does not require any DNA ligation and is very rapid. It allows a single gene or region on a cosmid to be replaced by a bi-functional selectable marker (having both an E. coli and an A. fumigatus marker). - [Read A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol]

Adapting Cells to a Serum-Free Environment Protocol - https://fscimage.fishersci.com/webimages_FSC/downloads/HyClone_Protocol_8.pdf

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

There are many ways to adapt cell lines to serum-free media. Five methods are presented that are designed for adapting hybridomas to a protein-free medium. These protocols may require some modifications for your particular cell line and conditions. - [Read Adapting Cells to a Serum-Free Environment Protocol]

Breaking the Tips of Embryonic Stem (ES) Cell Injection Needles Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4417

Category: Stem Cell Protocols: Stem Cell Injection Protocols

Injection needles require sharp tips that allow the blastocyst to be penetrated easily. This protocol describes a method of creating sharp injection needle tips. - [Read Breaking the Tips of Embryonic Stem (ES) Cell Injection Needles Protocol]

Forward Genetics in Arabidopsis: Finding Mutations that Cause Particular Phenotypes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/26/pdb.top1

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]

Glufosinate Ammonium Selection of Transformed Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4670

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol describes the use of glufosinate ammonium to select transformed Arabidopsis plants. The major advantage of glufosinate ammonium selection is that it can be performed on plants growing in soil and does not require the use of sterile techniques. - [Read Glufosinate Ammonium Selection of Transformed Arabidopsis Protocol]

Growing Adherent Cells Directly on Tissue Dishes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4290

Category: Cell Culture Protocols

For low-resolution work, cells to be used for staining can be grown directly on regular tissue-culture dishes. It is a convenient method that does not require much preparatory work. - [Read Growing Adherent Cells Directly on Tissue Dishes Protocol]

Growth of S. cerevisiae and Preparation of DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3298

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes methods for isolation of DNA from a strain of S. cerevisiae carrying a recombinant YAC. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. The method is suitable for preparing DNA that will be used for agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, PCR, or other methods that do not require intact high-molecular-weight DNA. - [Read Growth of S. cerevisiae and Preparation of DNA Protocol]

Immunofluorescent Staining for the Laser Microdissection of Individual Cells Protocol - http://www.natureprotocols.com/2006/08/03/immunofluorescent_staining_for.php

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Useful techniques to circumvent disruption of tissue structure in the analysis of gene expression are LCM and LDM. While they require specialized microscopes and systems, they are similar in that freshly-cut frozen tissue sections can be microdissected using either a general histological stain (like H&E) or by staining with fluorescently conjugated antibodies. The LCM system by Arcturus involves... - [Read Immunofluorescent Staining for the Laser Microdissection of Individual Cells Protocol]

Immunoprecipitation / Kinase Assay - http://www.upstate.com/misc/protocols.q.prot.e.ipkinaseassay/Immunoprecipitation+%2F+Kinase+Assay

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Immunoprecipitation / Kinase Assay Protocol. Upstate. Protocol applicable only to kinases whose activity is not altered by cell lysis or immunoprecipitation procedures, and do not require soluble cofactors for activity. - [Read Immunoprecipitation / Kinase Assay]

Preparation of Endothelial Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E538DDD4A302A0DF2CDDD9363072C1CA&objectid=6673B460F50B90057960CE47ABB708A4

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Endothelial cells, which line blood vessels, can be prepared from a variety of tissues. They are frequently prepared from the umbilical vein, which is relatively easy to obtain. The procedure is clearly described and provides a large population of highly purified endothelial cells. There are also methods for obtaining endothelial cells from other tissues such as fat, skin, and mucosa. These methods require special care and generate smaller populations of cells. - [Read Preparation of Endothelial Cells Protocol]

Protocol for Stable Lines - http://genetics.med.harvard.edu/~cepko/protocol/mike/S4.html

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

This protocol is optimized for erythroid cells, other cell types may require modifications. - [Read Protocol for Stable Lines]

Purification of RNA from Plant Tissue Using the Concert Plant Reagent - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4106

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

This protocol is not phenol-based, but does require the addition of chloroform. The Concert Plant Reagent is intended for the isolation of RNA from a wide variety of plant tissues including blue spruce needles, potato tuber etc. - [Read Purification of RNA from Plant Tissue Using the Concert Plant Reagent]

SCE protocol - http://www.jax.org/staff/mills/protocols/sce_protocol.pdf

Category: Cytogenetics Protocols

SCE protocol. Protocol for the use for wildtype MEFs. It may require some adjustments for other genotypes or cell types. - [Read SCE protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

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

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cytogenetics Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. Another inexpensive and easy-to-use alternative is to grow cells in a culture dish with a glass bottom. Such dishes are suitable for microinjection experiments. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Studying Mitosis in Cultured Mammalian Cells Prtocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: Mitosis Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. Another inexpensive and easy-to-use alternative is to grow cells in a culture dish with a glass bottom. Such dishes are suitable for microinjection experiments. - [Read Studying Mitosis in Cultured Mammalian Cells Prtocol]

The Cell Growth Environment - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_6.html#Environment

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

Cultured cells require a sterile environment and a supply of nutrients for growth. Basic Constituents of media. Sigma - [Read The Cell Growth Environment]

Articles (1-6 of 6)

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

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.

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose gels.

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

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.

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