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We narrowed to 1,023 results for: Tes

Showing: 821 - 840 of 1023 results
  1. 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...
  2. 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...
  3. 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...
  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. 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...
  7. 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...
  8. 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...
  9. 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...
  10. 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...
  11. 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 (Figure 1). 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 ...
  12. 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...
  13. 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 (Figure 1). 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 ...
  14. 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...
  15. 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...
  16. 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...
  17. 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...
  18. 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...
Showing: 821 - 840 of 1023 results