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We narrowed to 44 results for: NTS

Showing: 21 - 40 of 44 results

  1. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...equipment, reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other ...bath Vortex Stir bar Magnetic stir plate pH meter Reagents HEK293 cells Recombinant antibody plasmid DNA ...Pluronic-F68 40 mL 200 mM Glutagro Do not add selective reagents Store at 4 °C until use. We suggest preparing ...

  2. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...protein expression studies. Moreover, repeating experiments in a stable cell line, as opposed to transiently-transfected...incubator Pipet controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-...lines. It is not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles....may also exhibit varying numbers of integration events. In other words, if an MOI >1 was used, some cells...

  3. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...equipment, reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other ...optional) Scale Microsoft Excel or similar software Reagents Purified antigen Purified recombinant antibody...Tween-20, VWR 0777-1L Foil Before Starting Warm reagents to room temperature. Procedure Section 1: Prepare...

  4. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...population that varies in the number of integration events and the site(s) of proviral integration across ...Microscope Hemocytometer or other cell counter Reagents Polyclonal stable cell pool DMEM high glucose,...using selective medium throughout subsequent experiments. When selective medium is used, it may have additional... that the use of selective medium throughout experiments be determined empirically, and that the cell ...

  5. Lentivirus ddPCR Titration

    Type
    Protocol
    ..., where TU is transducing units and generally represents the number of infectious viral particles. Users... Mini Centrifuge, Thermo Scientific, 10199-452 Reagents GeneJet Genomic DNA Purification Kit, Thermo Fisher...equipment with 10% bleach prior to use and keeping all reagents and samples on ice or pre-chilled 96-well freezer...during use. To reduce the risk of contaminating reagents we recommend making small aliquots of master mixes...

  6. Water Bath Protocol

    Type
    Protocol
    ...Disinfectant Thermometer Water bath weights and floats Reagents None needed Procedure Ensure that the water bath...prevent growth of bacteria or fungi. There are disinfectants designed to be used in water baths with instructions...clean out the water bath even if you are using disinfectants as described above in step 3. You will also ...

  7. Ligation Independent Cloning

    Type
    Protocol
    ...overhangs to form a stable association between fragments, allowing for transformation without ligation....Vector In this example, the vector is cut at two points by the enzyme BsaI, removing the stuffer sequence... A typical T4 Pol reaction is shown. Mix the components (add the T4 last) and incubate at room temperature...

  8. Video Library

    Type
    Protocol
    ...purification allows you to isolate and purify DNA fragments based on size. Following gel electrophoresis, ...and Australia, and advice for current graduate students considering their future careers. Eric J. Perkins...Business Team, his hobbies, and advice for grauated students pursuing non-academic careers....

  9. Fluorescence Titering Assay

    Type
    Protocol
    ...waste container Inverted fluorescence microscope Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-... It is not recommended to subject lentiviral supernatants to multiple freeze-thaw cycles. Procedure Seed...you risk counting cells with multiple integration events leading to underestimation of the true titer. Calculate...

  11. Pouring LB Agar Plates

    Type
    Protocol
    ...works. Equipment Autoclave Water bath Pipetman Reagents 37 g pre-mixed powder consisting of: 5.0 g yeast...bottle with your initials, the date, and the bottle contents. This will clear up any confusion later if your...recommended temperature should kill most potential contaminants, certain types of spores will stubbornly survive...

  12. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...equipment, reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other ... Gel imaging system ImageJ or similar software Reagents Purified recombinant antibody, 0.9–1.1 mg/mL SimplyBlue...

  14. Western Blot

    Type
    Protocol
    ...equipment, reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other ...Platform shaker Cold room Gel imager -80 °C freezer Reagents 1X PBS Lysis buffer e.g., RIPA lysis buffer Microcentrifuge...

  15. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Magnetic stir bar Sonicator Ear protection Vortex Reagents Adherent HEK293T cells (ideally AAV-293T clones...Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other transfection reagents may be used in this protocol, but their conditions... with PEI and allow for the addition of other components, such as sorbitol and sodium bicarbonate, sorbitol...

  16. Protocol - How to Streak a Plate

    Type
    Protocol
    ...(or other small flame source) Incubator Marker Reagents LB agar plate (with appropriate antibiotic) Bacterial...DNA or use the individual colonies for other experiments....

  17. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...burner (or other small flame source) Incubator Reagents 6 cm diameter LB/agar plates without antibiotic...will work for this assay, and the above result represents a single experiment. For publishable data, the...

  18. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...purification allows you to isolate and purify DNA fragments based on size. The procedure starts with standard...Gel Electrophoresis Protocol with the following amendments: Notes: Gel purification is most efficient with...

  19. General Transfection

    Type
    Protocol
    ...different cell lines and different transfection reagents. Workflow Timeline Day 0: Seed HEK293T cells (...incubator Pipet controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-...

  20. Lentivirus Production

    Type
    Protocol
    ...alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for a variety...container pH meter Stir plate Magnetic Stir Bar Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-...
Showing: 21 - 40 of 44 results