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Showing: 961 - 1000 of 1038 results
  1. Fluorescence Titering Assay

    Type
    Protocol
    ...container Inverted fluorescence microscope Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5...each well of the 6-well dish. Incubate the cells overnight. If using freshly collected virus, filter through...If you anticipate that your viral stock will be higher titer, consider additional dilutions. Dilution ...
  2. Immunocytochemistry

    Type
    Protocol
    ...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...mg/mL DAPI solution to 100 µL PBS. Protect from light. Prepare a 300 nM DAPI working solution by diluting...DAPI stock solution into 5 mL PBS. Protect from light. Procedure Section 1: Seeding cells Place a sterile...Pro-Tip The plate can be wrapped in foil to block light. Remove the secondary antibody and dispose of it...
  3. Immunocytochemistry

    Type
    Protocol
    ...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...mg/mL DAPI solution to 100 µL PBS. Protect from light. Prepare a 300 nM DAPI working solution by diluting...DAPI stock solution into 5 mL PBS. Protect from light. Procedure Section 1: Seeding cells Place a sterile...Pro-Tip The plate can be wrapped in foil to block light. Remove the secondary antibody and dispose of it...
  4. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ...transfection into mammalian cells is fairly straightforward and the resultant cells can either express ...
  5. Colony Formation Titering Assay

    Type
    Protocol
    ...controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat inactivated FBS and 5...colonies for at least 2 of the dilutions. Pro-Tip The higher dilution wells tend to provide more accurate titers...dilutions, and single colonies are visible in the higher dilutions. Last reviewed on: August 31, 2023...
  6. Colony Formation Titering Assay

    Type
    Protocol
    ...controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat inactivated FBS and 5...colonies for at least 2 of the dilutions. Pro-Tip The higher dilution wells tend to provide more accurate titers...dilutions, and single colonies are visible in the higher dilutions. Last reviewed on: August 31, 2023...
  7. Pipetting Protocol

    Type
    Protocol
    ...to begin using the pipette. To the right is a photo of how you might hold a pipette. Without moving past... P1000 pipette shows the volume adjustment ring right below the plunger. This ring changes the pipette...P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume P2 0.2 to 2 µL P10 1 to...
  8. Pipetting Protocol

    Type
    Protocol
    ...to begin using the pipette. To the right is a photo of how you might hold a pipette. Without moving past... P1000 pipette shows the volume adjustment ring right below the plunger. This ring changes the pipette...P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume P2 0.2 to 2 µL P10 1 to...
  9. Antibodies 101: Isotypes

    Type
    Blog Post
    ... an antibody labeled with Fc, Fab, heavy chain, light chain, constant region, and variable region. ...
  10. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...purified AAV. Option #3 Puncture the QuickSeal tube slightly below the 60–40% interface with an 18 ga needle... (1999): 973-985 (Link opens in a new window) . Right panel: cartoon indicating the position of the needle...harvesting of the purified AAV using option #1. Right image adapted by permission from Macmillan Publishers... viral vectors," (Link opens in a new window) copyright (2006). Figure 3: Example of an SDS-PAGE gel of...
  11. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...purified AAV. Option #3 Puncture the QuickSeal tube slightly below the 60–40% interface with an 18 ga needle... (1999): 973-985 (Link opens in a new window) . Right panel: cartoon indicating the position of the needle...harvesting of the purified AAV using option #1. Right image adapted by permission from Macmillan Publishers... viral vectors," (Link opens in a new window) copyright (2006). Figure 3: Example of an SDS-PAGE gel of...
  12. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...limitation of using loxP sites is the inability to tightly control which loxP sites recombine if more than...
  13. Working with Nuclear Receptors

    Type
    Blog Post
    ...The coregulator ‘core’ complex is composed of a tightly-bound invariant group of proteins, whereas the ...
  14. Lentivirus ddPCR Titration

    Type
    Protocol
    ...viral particles. Users may need to run lower or higher dilutions depending on their particular sample....tissue culture treated dish, VWR, 29442-042 DMEM, high glucose, pyruvate, Corning, 10-013-CV Heat-inactivated... Reagent Preparation DMEM Complete: 500 mL DMEM, high glucose, pyruvate 55 mL heat inactivated premium...increases by a factor of ~2 as you progress from the higher dilutions to the lower dilutions (blue). As shown...
  15. Lentivirus ddPCR Titration

    Type
    Protocol
    ...viral particles. Users may need to run lower or higher dilutions depending on their particular sample....tissue culture treated dish, VWR, 29442-042 DMEM, high glucose, pyruvate, Corning, 10-013-CV Heat-inactivated... Reagent Preparation DMEM Complete: 500 mL DMEM, high glucose, pyruvate 55 mL heat inactivated premium...increases by a factor of ~2 as you progress from the higher dilutions to the lower dilutions (blue). As shown...
  16. DNA Quantification

    Type
    Protocol
    ...spectrophotometer uses the absorbance/transmission of light through a liquid to determine the concentration ...liquid. Molecules absorb different wavelengths of light to varying degrees and most have a specific wavelength... absorbance, you need to know the wavelength of light that your substance maximally absorbs. In the case...
  17. DNA Quantification

    Type
    Protocol
    ...spectrophotometer uses the absorbance/transmission of light through a liquid to determine the concentration ...liquid. Molecules absorb different wavelengths of light to varying degrees and most have a specific wavelength... absorbance, you need to know the wavelength of light that your substance maximally absorbs. In the case...
  18. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...constructs, ADAR2DD(E488Q/T375G) (REPAIRv2) had the highest on-target efficiency and the lowest amount of off-target...
  19. An Introduction to Adenovirus

    Type
    Blog Post
    ...vectors all have a genomic capacity that is much higher than other viral vectors (tens of kilobases; compared...
Showing: 961 - 1000 of 1038 results