convenient Protocols

Category: Fungi Protocols: Fungi Genetics Protocols

Protocol describes a safe and convenient method of extracting DNA from Coccidioides immitis fungi in which the culture is killed by steaming, allowing removal from the containment facilities, as soon as possible. The method was first developed with the non-pathogen Neurospora crassa, has worked well for both C. immitis and H. capsulatum, and should be useful for extracting DNA from any pathogenic fungus. - [Read A Safe Method of Extracting DNA from Coccidioides immitis Protocol]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Absorbance assay at 280 nm.  This method is just as convenient as for absorbance at 280 nm.  It may be preferred if there is excessive contamination by nucleic acids, since nucleic acids absorb very little radiation at 205 nm.  Setting the wavelength is a bit tricky since 205 nm is right on the shoulder of the protein peak. - [Read Absorbance Assay 205 nm]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Absorbance assays are fast and convenient, since no additional reagents or incubations are required.  No protein standard need be prepared.  The assay does not consume the protein.  The relationship of absorbance to protein concentration is linear.  Because different proteins and nucleic acids have widely varying absorption characteristics there may be considerable error, especially for unknowns or protein mixtures. - [Read Absorbance Assay 280 nm]

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

The most convenient and commonly used method to visualize DNA in agarose gels is staining with the fluorescent dye ethidium bromide.  Ethidium bromide can be used to detect both singleand double-stranded nucleic acids (both DNA and RNA).  However, the affinity of the dye for single-stranded nucleic acid is relatively low and the fluorescent yield is comparatively poor. - [Read Detection of DNA in Agarose Gels Protocol]

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]

Category: Histology Protocols: Laser Capture and Microdissection

Laser Capture Microdissection of Living in vitro Cells. This PDF describes a precise, rapid and convenient Laser Capture Microdissection (LCM) method for the positive selection of living adherent cells and the successful subsequent re-cultivation of homogenous sub-populations. Arcturus. - [Read Laser Capture Microdissection of Living in vitro Cells PDF]

Category: C. elegans Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. - [Read Live Imaging of Caenorhabditis elegans: Examples]

Category: Tetrahymena Protozoa Protocols

Long-term storage of unfrozen Tetrahymena is convenient when a laboratory is not using the cells frequently; however, cells maintained under these conditions often take a couple of days of incubation in culture medium to recover. - [Read Long-Term Storage of Unfrozen Tetrahymena Cultures in Soybean Medium Protocol]

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]

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

Protocols utilize T hybridomas to detect expression of peptide-MHC complexes, since these cells provide the most convenient, consistent, and flexible T cell readout systems for these purposes. If desired, antigen-specific T cell clones can be used in lieu of T hybridoma cells, but T cell clones often give poorer responses than T hybridomas to fixed APCs due to fixation-induced loss of costimulator function. - [Read Presenting Exogenous Antigen to T Cells Protocol]

Category: Microscopy Protocols: In Vivo Imaging Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]

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

Protocol describes a convenient, although imprecise, means of enriching T cells through removal of accessory and B cells; use of nylon wool is preferred if both of the latter subsets are to be removed, while Sephadex is used when the goal is primarily to remove accessory cells. - [Read T Cell Enrichment By Nonadherence to Nylon Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase assay]

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase Assay Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable. - [Read Transient Transfection Into 293T Cells Protocol]