minimal Protocols

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]

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

Annexin V, belonging to a recently discovered family of proteins, the annexins, with anticoagulant properties has proven to be a useful tool in detecting apoptotic cells since it preferentially binds to negatively charged phospholipids like PS in the presence of Ca2+ and shows minimal binding to phosphatidylcholine and sphingomyeline. Changes in PS asymmetry, which is analyzed by measuring Annexin V binding to the cell membrane, were detected before morphological changes associated with... - [Read Annexin V Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

4 strains of E. coli are used in these studies: JM101 for M13 infection and isolation, XL1BMRF'for M13 or pUC-based DNA transformation, and ED8767 for cosmid DNA transformation. To maintain their respective F' episomes necessary for M13 viral infection, JM101 is streaked onto a M9 minimal media plate and XL1BMRF' is streaked onto an LB plate containing tetracycline. ED8767 is streaked onto an LB plate. These plates are incubated at 37degC overnight. For each strain, 3 ml. of appropriate liquid.. - [Read Bacterial Cell Maintenance Protocol]

Category: Plant Biology Protocols: Plant Tissue Culture

Includes: Chu (N6) basal salt mixture DKW/Juglans basal salt mixture Gamborg's B-5 basal salt mixture Gamborg's B-5 basal salt mixture with minimal organics Hoagland's No. 2 basal salt mixture McCown's woody plant basal salt mixture Murashige and Skoog basal salt mixture (MS) Quoirin and Lepoivre basal salt mixture Schenk and Hildebrandt basal salt mixture White's basal salt mixture - [Read Classic Plant Media]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Specimen chambers have had many designs published over the years describing systems that offer excellent optical properties while allowing specimens to be maintained for varying amounts of time. Ranging in complexity from the simple preparation of a sealed coverslip on a microscope slide to sophisticated perfusion chambers that enable tight control of virtually all environmental variables culture chambers are designed to to allow living specimens to be observed with minimal invasion at high res. - [Read Culture Chambers for Live-Cell Imaging]

Category: Fluorescent Dyes Protocols

Cyanine dye reagents are useful as fluorescent labels for proteins. This protocol has been designed to label the thiol group on cysteine using Cy3 or Cy5 minimal maleimide labeling dyes. - [Read Cyanine Dye (Maleimide) Protein Labeling Protocol]

Category: Protein Protocols: Protein Expression Protocols

Expression of protein in minimal media using E. coli cells for expression. Dr. Chen, Biochem. & Mol. Biology, University College. London - [Read Expression of protein in minimal media]

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

How to prepare M9 minimal media and grow in M9 media. Chazin Lab , Vanderbilt Univ. - [Read Expression Protocol in M9 Minimal Media via T7 Promoter]

Category: Bacterial Cell Culture Protocols: Bacterial Cell Culture Media Protocols

This protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 minimal media begins with preparing a 5x stock solution of M9 salts. Generally, M9 salts contain a nitrogen source in the form of NH4Cl. Since we want to add a labeled nitrogen source, our 5x salts are prepared minus NH4Cl. Standard 5 X M9 Minimal Media salts minus nitrogen source For 1L 5xM9 salts: - [Read Expression Protocol in M9 Minimal Media via T7 Promoter]

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

Protocol for fission yeast media. Includes: YES (yeast extract with supplements) for rich complete media; YSO, for modified rich media; EMM (Edinburgh minimal medium) for minimal selective media; stock solutions to make up minimal media; MB, a very stringent minimal for transformation; Sporulation media; ME (malt extract); SPAS; EMMGlut. - [Read Fission Yeast Media Protocol]

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for lactose RNAi plates: minimal lactose media. - [Read Lactose RNAi Plates: Minimal Lactose Media Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Cell Culture Media Protocols

Protocol for M9 minimal media (1000 ml). - [Read M9 Minimal Media (1000 ml) Protocol]

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

Protocol for media for culture of Aspergillus nidulans. Includes minimal and complete media. - [Read Media for Culture of Aspergillus nidulans Protocol]

Category: Fungi Protocols: Aspergillus Protocols

Protocol for media for culture of Aspergillus nidulans. Includes: Minimal medium; Complete media. - [Read Media for Culture of Aspergillus nidulans 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 Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

In an attempt to accurately measure DNA content with simultaneous preservation of cell surface markers, we have utilized gentle ethanol treatment techniques, which permeablize cells with minimal loss of surface antigen expression and antibody-antigen association. For some cell types, the presence of apoptotic cells based on reduced DNA content can also be detected. One such technique employs the addition of ethanol to cells previously resuspended in high concentrations of fetal bovine serum... - [Read Simultaneous Analysis of DNA Content and Surface Immunophenotype 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 Culture Protocols

Protocol describes static culture of postimplantation embryos, an alternative to the roller method. The static method is best suited to 6.0 to 7.0 days post coitum (dpc) embryos followed for 24 hours (7.0 dpc embryos) to 48 hours (6.0 dpc embryos) of development. It allows repetitive real-time observation with minimal handling of the embryo. It is especially useful if single or small groups of embryos need to be distinguished from each other. - [Read Static Culture of Postimplantation Embryos Protocol]

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

Vogel's 50X salts as given in the Microbial Genetics Bulletin 13:42-43, 1956. In 750 ml. distilled water, dissolve successively with stirring at room temperature. - [Read Vogel's Minimal Media Protocol]

Category: Fungi Protocols

Protocol for Vogels medium N (minimal). - [Read Vogels Medium N (Minimal) Protocol]