medium Protocols

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

Boris Greber. Important rules for ES cell culture, Signs of differentiation, General tips, ES medium, Thawing ES cells, Freezing, Passaging. - [Read Mouse embryonic stem (ES) cell culture - basic procedures]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Great Procedures. rules for ES cell culture, differentiation of ES cells, ES medium, Thawing, Passaging, Freezing. Mouse embryonic stem (ES) cell culture - basic procedures. Max Planck Institute for Molecular Genetics. Boris Greber. - [Read Mouse embryonic stem (ES) cell culture - basic procedures PDF]

Category: Plant Biology Protocols: Plant Tissue Culture

A recipe for MS plant medium. Details. Details of this protocol, MS Plant Tissue Culture Medium MS Plant Tissue Culture Medium Protocol - [Read MS Plant Tissue Culture Medium Protocol]

Category: Fungi Protocols: Neurospora Protocols

Protocols for Neurospora methods. Includes: Standard strains; Crosses; Minimal medium; Color coding of media; Agar substrate for manipulation and isolation; Stock solutions of supplements; Mating-type tests; Preservation of stocks by silica gel; Cleaning of glassware; Control of mites. - [Read Neurospora Methods Protocols]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Remove medium from culture container, rinse with PBS, expose cells to trypsin, incubate, stop trypsin activity by adding serum containing medium, dissociate cells, dilute and return to incubator. - [Read Passage of Adherent Cell Lines (subculture).]

Category: Molecular Biology Protocols: Phage Protocols and Methods

Preparation of liquid medium for phage growth. Rob Phillips Group: Physical Biology of the Cell - [Read Phage Media Preparation]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Zygotes can be identified by their unique morphology. They can be easily separated away from nonmated cells using a micromanipulator. This method provides an alternative to the selection of diploid cells on a medium that prevents the growth of haploid parent cells. - [Read Picking Zygotes Protocol]

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected cells into the surrounding medium. The filamentous particles are concentrated by precipitation from a high-ionic-strength buffer with polyethylene glycol. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol. This protocol is generally used to prepare single-stranded DNA from a small number of M13 isolates. - [Read Preparation of Single-stranded Bacteriophage M13 DNA Protocol]

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]

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]

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts 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]

Category: Fungi Protocols

Protocol for bird medium. This medium was designed to circumvent some problems that arise in the use of Medium N. - [Read Protocol for Bird Medium: Alternative to Vogel Medium]

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]

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]

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]

Category: Cell Biology and Cell Culture

Protocol for the recovery of a highly viable fraction for use in fertilization. Protocol uses an iodinated density gradient medium to adjust the density of the raw ejaculate to approx 1.170 g/ml by mixing with a high density medium (ρ >1.26 g/ml) and loading it beneath a discontinuous gradient. - [Read Purification of viable bovine spermatozoa of normal morphology.]

Category: DNA Protocols: cDNA Library Protocols

A cDNA library constructed in a plasmid expression vector of the pUC, pUR, or pEX series is plated on agar medium and then replicated onto filters, which are transferred to plates containing IPTG. After 2-4 hours of induction, the colonies are lysed with chloroform and then screened with appropriate antibodies. - [Read Screening Expression Libraries Constructed in Plasmid Vectors Protocol]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Protocol for sequential adaptation to HyQ© SFX-insect™ medium. - [Read Sequential Adaptation to HyQ© SFX-Insect™ Medium Protocol]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Protocol for spore germination. This procedure is typically used for the isolation and preparation of spores from a diploid strain heterozygous for a marked disruption (e.g., yfg1::his3+) Inoculation of the spore population into minimal medium lacking the nutritional supplement corresponding to the disruption marker (e.g., minimal medium lacking histidine) allows only the disruption spores to germinate. - [Read Spore Germination Protocol]

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

Protocol for sterilization of plant culture media. Includes: Minimum Autoclaving Time for Plant Tissue Culture Medium. - [Read Sterilization of Plant Culture Media Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Protocol describes a method for storing bacterial cultures that are growing on solid media. It is useful for storing copies of cDNA. - [Read Storage of Bacterial Cultures Growing on Solid Medium Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Phage are streaked onto a medium to obtain an independent isolate prior to preparing a new lysate. This is done to reduce the likelihood of working with lysates which have become contaminated, and/or have accumulated mutations. - [Read Streaking Lambda Phages Protocol]

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

Some yeast strains are unstable (e. g., small YAC-bearing strains) and need to be repurified by streaking on an agar plate and then verifying the genetic content of the isolated colony before proceeding. In cases where the strain is unstable, plan to streak the cells onto the selective medium to retain the desired stock, (however, most strains can be streaked onto the complete medium, YPD). - [Read Streaking Yeast Stocks Protocols]

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

Protocol for TC-100 insect medium. - [Read TC-100 Insect Medium Protocol]

Category: Mouse Protocols: Mouse Cell Culture Protocols

The liver of a rat is cannulated and perfused in situ with buffer, following which it is excised and perfused in a closed system with a collagenase solution. After a period of time the liver begins to break up, at which point it is transferred to a measuring cylinder and culture medium is added. It is then gently agitated to cause the release of cells which are subsequently filtered and allowed to settle out. The parenchymal and non-parenchymal cells form two distinct layers which can be separat - [Read The Isolation and Culture of Rat Hepatic Cells Protocol]