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Addgene

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We narrowed to 44 results for: Tes

Showing: 21 - 40 of 44 results

  1. Lentivirus Production

    Type
    Protocol
    ...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ... plate in DMEM Complete in 10 cm tissue culture plates. Incubate the cells at 37 °C, 5% CO 2 for ~20 h...the table below for a possible range of ratios to test: Ratio of DNA:PEI μg of DNA μL of 1 mg/mL PEI 1:...Centrifuge the viral supernatant at 2100 rcf for 5 minutes to pellet any packaging cells that were collected...

  2. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Biosafety Cabinet 4 °C Refrigerator Micropipettes and tips Pipettor and pipettes Benchtop centrifuge compatible...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...Reagent Preparation instructions. Wipe down all pipettes and equipment with 10% bleach prior to use. Reagent...

  3. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...transiently-transfected cells, increases reproducibility, as it eliminates the variation associated with repeated transient...and FBS lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers to find ... expanded, prepare cell stocks and/or harvest to test for protein expression. Typically, cells transduced...levels of the transgene may have reduced growth rates, especially if that transgene is toxic. Eventually...

  4. AAV ddPCR Titration

    Type
    Protocol
    ...primers, and probe on ice before use. Wipe down all pipettes and surfaces with 10% bleach. Safety Warnings ...PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 48-well dilution plate, Bio-... 1X PCR buffer (1:25,600,000) Use multichannel pipettes for the dilution series. For dilutions 1–5, use...reduce NTC values, we recommend wiping down all pipettes and equipment with 10% bleach prior to use and...

  5. AAV Production in HEK293 Cells

    Type
    Protocol
    ...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ...inactivated in the lab by heating to 56 °C for 30 minutes. 0.45 μm polyethersulfone (PES) filter system, ...and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently tap the sides of the... and sonicate 5 x 1 sec pulses with at least 5 minutes on ice between each pulse, 50% amplitude. Return...

  6. Lentivirus ddPCR Titration

    Type
    Protocol
    ...mixes and samples on ice before use. Wipe down all pipettes and surfaces with 10% bleach. Safety Warnings ...PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable foil heat seal, Bio-Rad...Cell Harvest Gently aspirate media from the 6-well plates. Be sure to use a new aspirating pipette for each...and Troubleshooting We recommend wiping down all pipettes and equipment with 10% bleach prior to use and...

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

    Type
    Protocol
    ...with standard agarose gel electrophoresis, which separates DNA by their length in base pairs. Following electrophoresis...Electrophoresis Protocol with the following amendments: Notes: Gel purification is most efficient with lower %...blade, slice the desired DNA fragment from the gel. Notes: To protect the UV box, it is a good idea to place...

  8. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...commonly used to culture bacteria in the lab. LB agar plates are frequently used to isolate individual (clonal...characterized by a cloudy haze in the media (see right). Notes: Some protocols require bacteria to be in the log...on your plasmid. If the bacteria on your LB agar plates are not fresh, you should streak your bacteria ...

  10. General Transfection

    Type
    Protocol
    ...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ... plate in DMEM complete in 10 cm tissue culture plates. Incubate the cells at 37 °C, 5% CO 2 for ~20 h...the table below for a possible range of ratios to test: Ratio of DNA:PEI Amount of DNA (μg) Volume of 1...

  11. Gibson Assembly Protocol

    Type
    Protocol
    ...Gibson Cloning? No need for specific restriction sites. Join almost any two fragments regardless of sequence...unique function for the reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing back ...other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed...

  12. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...ultracentrifugation. The arrow indicates the 60–40% interface. The vertical black line indicates the location of the...fractions once you have a good idea when the virus elutes from the gradients. In a biosafety cabinet, carefully...

  13. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...tubes, VWR 76289-498 Before Starting Wipe down all pipettes and equipment with 10% bleach prior to use. Warm...Spectrophotometer using the A280 IgG setting. Combine all eluates with measurable protein. Determine the protein ...

  14. Western Blot

    Type
    Protocol
    ...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...January 24, 2022 Workflow Timeline Day 1: Prepare lysates, run SDS-PAGE, transfer, block, incubate with primary...channel pipette Pipette controller Pipette tips and pipettes Spectrophotometer Heat block Mini gel tank chamber...

  15. Water Bath Protocol

    Type
    Protocol
    ...on the surfaces of the water bath as the water evaporates from the tub. Disinfectant may be added to prevent...heat up quicker. Pro-Tip Set up the water bath 30 minutes to 1 hour before you need to use it to allow the...

  16. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ... 0.5-1μL T4 DNA Ligase H 2 O to a total of 10μL Notes: If the DNA concentrations are low such that you...troubleshooting failed ligations. The following table indicates the various controls: Control Ligase Interpretation...

  17. Immunocytochemistry

    Type
    Protocol
    ...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment... Equipment Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope...

  18. Colony Formation Titering Assay

    Type
    Protocol
    ...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ...solution through a 0.22 μm filter to remove any precipitates. Stain each well with 1 mL of 0.1% crystal violet...

  19. Protocol - How to Design Primers

    Type
    Protocol
    ...also needs to avoid primer-primer annealing which creates primer dimers and disrupts the amplification process...
Showing: 21 - 40 of 44 results