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Preparation of Genomic DNA from Yeast Using Glass Beads Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4151

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

Protocol describes the preparation of genomic DNA from a 5-ml liquid culture of yeast cells. - [Read Preparation of Genomic DNA from Yeast Using Glass Beads Protocol]

Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4483

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

MEF feeders are prepared weekly to provide a substrate for undifferentiated embryonic stem (ES) cells. Primary MEF cells are thawed, established in culture, treated with mitomycin C to halt their proliferation so they cannot overgrow the ES cultures, and then replated onto dishes convenient for ES cell culture. This protocol can also be used to prepare feeder cells from STO fibroblast cell lines. - [Read Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol]

Preparing Stocks of Bacteriophage {lambda} by Small-scale Liquid Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3971

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

High-titer stocks of bacteriophage {lambda} are easily prepared by infecting small-scale bacterial cultures. - [Read Preparing Stocks of Bacteriophage {lambda} by Small-scale Liquid Culture Protocol]

Primary Culture of Rat Cerebral Astrocytes Protocol - http://josm.geneseo.edu/theses/Weiss-02.pdf

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Protocol for primary culture of rat cerebral astrocytes. - [Read Primary Culture of Rat Cerebral Astrocytes Protocol]

Procedures for In Vitro Production of Bovine Embryos - http://www.animal.ufl.edu/hansen/IVF/default.htm

Category: Developmental Biology: Embryology Protocols

Procedures for in vitro production of bovine embryos. Includes: Collection of Ovaries; Oocyte Collection; Preparation of IVF Medium; In Vitro Fertilization; Culture of Embryos. - [Read Procedures for In Vitro Production of Bovine Embryos]

Process Protocol for Hepatocyte Harvest - http://www.biolifesolutions.com/protocols/Hepatocyte_Protocol.pdf

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

Process Protocol for Hepatocyte Harvest ... Re-suspend as necessary in culture media (for cell culture) or in CryoStor ... - [Read Process Protocol for Hepatocyte Harvest]

Proliferative Assays for T Cell Function Protocols - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6634679E96E622003E15187F105306&objectid=6674B7F5E219FD4021F448C7D13AF27A

Category: Immunology: T Cell Protocols

Protocols describe the conditions required to induce proliferation are described. Also describe the assay in which CD4·CD25·T cells are co-cultured with conventional T cells in order to assess their suppressive function. Will describe the culture conditions for the activation and expansion of CD4·CD25· cells. - [Read Proliferative Assays for T Cell Function Protocols]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4437

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA in tissue culture cells using either propidium iodide (PI) or DAPI staining followed by flow cytometry. PI can be excited at 488 nm by the argon-ion laser, the most commonly used laser in flow cytometry. Alternatively, DAPI is best excited by a high-power UV laser, which is less commonly available. - [Read Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry Protocol]

Protein Expression and Purification Protocol - http://sosnick.uchicago.edu/Protein_Express&Purify.html

Category: Recombinant Protein Protocols

From plasmid to protein using bacterial expression. Transform appropriate DNA plasmid, Make a starter culture for protein expression, bacteria culture for protein expression. Sosnick Group Chicago. - [Read Protein Expression and Purification Protocol]

Protocol for Primary Cultures of HUVECs - http://www.huvec.com/documents/Protocol-HUVECs.pdf

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Protocol for primary cultures of HUVECs. Includes: Buffers and reagents (Fetal calf serum, Phosphate Buffer Saline (PBS), Collagenase, Buffer for conservation and transport of umbilical cords, Culture medium); Cell culture. - [Read Protocol for Primary Cultures of HUVECs]

Protocol Primary Mammalian Endothelial Cells - http://www.amaxa.com/fileadmin/groups/marketing/Downloads/Protocols/Primary_cells/amaxa_OP_Basic_Endothelial_VPI-1001.pdf

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Primary mammalian endothelial cells protocol. This protocol is designed for primary endothelial cells isolated from various organs of mammals. Large and flat cells, often with large nuclei. Includes: Required reagents; DNA preparation and quality; Preparation of cells and cell culture; Important controls; Nucleofection protocol. - [Read Protocol Primary Mammalian Endothelial Cells]

Protocols for Insect Cell Culture - http://www.hyclone.com/library/insectprotocols.htm

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Includes protocols: Direct Adaptation to HyQ© SFX-Insect™ Medium; Sequential Adaptation to HyQ© SFX-Insect™ Medium; Adaptation of High Five™ Cells to HyQ© SFX-Insect™; Production of Recombinant Protein Using the Baculovirus Expression Vector System; Culturing Insect Cells and Production of Baculovirus Expressed Proteins in 2.8 L Fernbach Systems. Production of Autographa californica Multiple Capsid Nuclear Polyhedrosis Virus (rAcMNPV) Stock - [Read Protocols for Insect Cell Culture]

Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions - http://www.geron.com/PDF/scprotocols.pdf

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

Includes protocols: Mouse Embryonic Fibroblasts (MEF) Preparation; Harvesting MEFs; Cryopreservation of MEFs; Thawing and maintaining MEFs; Irradiating & Plating MEFs; Culture of Human ES cells with Matrigel® and Conditioned Medium; Preparation of Conditioned Medium (CM); Preparation of Matrigel® -coated plates; Passage of human ES cells on Matrigel®; Daily maintenance of feeder-free culture; Freezing Human ES Cells; Thawing Human ES cells; Formation of Embryoid Bodies; - [Read Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions]

Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio - http://www.fhcrc.org/science/labs/fero/Protocols/Hematopoietic.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Antibodies for Lineage Depletion; Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio]

Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells Protocol - http://www.fhcrc.org/science/labs/fero/Protocols/Hematopoietic.pdf

Category: Immunology

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells Protocol]

Quality Control Considerations for Cell Culture - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_9.html#Quality%2520Control

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

Quality is important in all aspects of tissue culture since the quality of materials used i.e. media and other reagents) will affect the quality of the cultures and products derived from them. The main areas of quality control that are of concern for tissue culture are: The quality of the reagents and materials; The provenance and integrity of the cell lines; The avoidance of microbial contamination. - [Read Quality Control Considerations for Cell Culture]

Quality Control Considerations for Cell Culture - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_9.html#Quality%20Control

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

Quality control considerations for cell culture. Includes: Provenance and Integrity of Cell Lines; Avoidance of Microbial Contamination; Environmental Monitoring; What to do in the event of contamination; - [Read Quality Control Considerations for Cell Culture]

Rapid Preparation of DNA from Cells in 96-Well Tissue Culture Dishes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4413

Category: Stem Cell Protocols

This protocol describes a method for rapid preparation of DNA from cells in 96-well tissue culture dishes. - [Read Rapid Preparation of DNA from Cells in 96-Well Tissue Culture Dishes Protocol]

Refeeding Marine Euplotids with Bacteria Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4468

Category: Laboratory Model Organisms Protocols

Feeding euplotids with algae can lead to asynchronous cell starvation and vastly different cell sizes within a culture. Asynchronous starvation also leads to different levels of mating competence. Furthermore, algal pigment remnants can interfere with many applications (e.g., fluorescence microscopy). - [Read Refeeding Marine Euplotids with Bacteria Protocol]

Removal of Yeast Contamination from Lymphoblast Cultures Protocol - http://humgen.wustl.edu/hdk_lab_manual/hcc/hcc8.html

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

This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when slightly shaken and lymphocytes will not cluster together as much as normal. If cultures are suspect, a drop of culture can be streaked on a YPD media plate to check for growth of yeast colonies, or a 5 ml sample can be taken to Barnes Diagnostic Center for identification of yeast strain. - [Read Removal of Yeast Contamination from Lymphoblast Cultures Protocol]

Root Transformation of Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4671

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Arabidopsis can be stably transformed using Agrobacterium tumefaciens-mediated transfer of T-DNA. We describe the generation of transgenic plants via root transformation in tissue culture, which can be useful for transforming sterile mutants. - [Read Root Transformation of Arabidopsis Protocol]

Serum Testing for Mammalian Cell Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4348

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

This protocol describes a method for testing of new serum lots prior to use in cell culture. Serum lots vary considerably in their ability to support cell growth, and some lots even contain toxic or growth-inhibitory compounds. It is advantageous to test serum lots and purchase in large volume, both for cost benefit. - [Read Serum Testing for Mammalian Cell Culture Protocol]

Setting Up Microdrop Cultures Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4353

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol describes how to set up microdrop cultures to produce embryos which can then be used for making chimeras. The microdrop culture should be set up several hours to 1 day before the experiment to permit temperature and gas equilibration. - [Read Setting Up Microdrop Cultures 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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