maintenance Protocols

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: General Research Protocols and Methods: Centrifuge Use and Centrifugation

Centrifuge Safety - High Speed Hazards. Safe Centrifuge Use , Care and Maintenance, and maintaining a log book for centrifuges. EHS Washington state Univ. - [Read Centrifuge Safety - High Speed Hazards]

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila growth conditions. Includes: MAINTENANCE; Semi-adherent cell lines; Adherent cell lines; SL2, S2C, S2*; DL2, DL1, S2R+, Kc167, Clone 8, S3; BG2. - [Read Drosophila Growth Conditions Protocol]

Category: Transfection Protocols: Stable Transfection Protocols

CHO Lec3.2.8.1 cells have 4 independent mutations in the N and O glycosylation pathways. When cultured with alpha-glucosidase I inhibitor N-butyl-deoxynojirimycin, glycoproteins produced in CHO Lec3.2.8.1 cells are completely susceptible to Endo H digestion. Endo H cleaves chitobiose, leaving a single N-linked N-acetylglucosamine per site which is ideal for maintenance of protein solubility and special carb-protein interactions, such as between the first N-acetyl glucosamine residue and tryp. - [Read Establishment of Stable Transfectant of CHO Lec Cells Protocol]

Category: General Research Protocols and Methods: Centrifuge Use and Centrifugation

Excellent Centrifuge Safety Use Manual. 30 pages. Training, Use, Maintenance, Records and Responsibility, Mechanical Breakdown, Emergency Procedure, Specific Safety Instructions Beckman Sorvall and Eppendorf. HSE UBC. - [Read Excellent Centrifuge Safety Use Manual PDF]

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

Information on Immortalized Mammary Epithelial Cells (IMEC). Includes: Background Information on IMEC; Culturing; Trypsinization. - [Read IMEC Maintenance]

Category: Tetrahymena Protozoa Protocols

Protocol provides a method for long-term storage of Tetrahymena cultures using maintenance culture medium. - [Read Long-Term Storage Storage of Unfrozen Tetrahymena Cultures in Maintenance Culture Medium Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on Luria-Bertani (LB) agar on or in LB broth containing Ampicillin (30 µg/ml) or Carbenicillin (50 µg/ml broth, 100 µg/ml agar). E. coli strains are usually preserved in stab agar or glycerol for mid-term storage and lyophilized for long-term storage. - [Read Maintenance of Probes in Bacteria Including Escherichia coli Protocol]

Category: Laboratory Model Organisms Protocols

Protocol describes the preparation of stocks of marine hypotrichs for long-term storage. Marine euplotids are maintained by serial passage of tube stocks. - [Read Maintenance of Stocks of Marine Hypotrichs Protocol]

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

This protocol has been established to maintain the efficient use of the AutoMACS cell sorter during periods of heavy use (e.g., three preparations of splenocytes from 16 spleens per week). Includes: Running the SAFE Clean Program; Clearing Debris. - [Read Maintenance of the AutoMACS Cell Sorter Protocol]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Method for Establishment and Maintenance of Leaf and Root Cell Cultures Arabidopsis thaliana. CL ENCINA1, M CONSTANTIN2 and J BOTELLA2. Plant Molecular Biology Reporter 19: 245–248, 2001. - [Read Method for Establishment and Maintenance of Leaf and Root Cell Cultures Arabidopsis thaliana]

Category: Cell Culture Protocols: Cell Lines Protocols

RAW 264.7 cells are a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice. For routine maintenance in culture (passage), cells are seeded at a confluence of approximately 10% (1 x 106 and 3 x 106 cells in 100-mm and 150-mm plates, respectively) and grown to a confluence of approximately 80%. This procedure requires the cells to be split every two days. - [Read Passage Procedure for RAW 264.7 Cells]

Category: Immunology: T Cell Protocols

Provides methods for the derivation of specific types of T cell clones: preparation and maintenance of alloreactive murine helper T (TH) lymphocyte and cytotoxic T lymphocyte (CTL) clones using the limiting dilution technique and derivation of TH clones reactive with soluble protein antigens including a method for the selection of either TH1 or TH2 lymphocyte subsets. - [Read Production of T Cell Clones Protocol]

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: Cell Culture Techniques

Tissue Culture Methods. Julie B. Wolf, Univ. Maryland. Protocols for Counting Cells, Preservation and storage of cell lines, maintenance of cell lines, media and growth, and safety guidelines for working with cells. - [Read Tissue Culture Methods]

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

High Efficiency Transformation of a yeast strain requiring maintenance of a plasmid. - [Read Two Hybrid System Transformation Protocol]