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Showing: 901 - 950 of 978 results

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ...PES. Using a PES filter will maximize titer . DMEM high glucose, Corning 10-013-CV DMEM, low glucose (1 ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5...dissolved. Pro-Tip This step is challenging due to the high viscosity of PEG. Stirring under medium heat will...precipitation. Precipitation of the viruses can proceed overnight at 4 °C if needed. Transfer the entire sample ... prep . The clarified supernatant can be kept overnight at 4 °C before proceeding with the purification...

  10. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...Inoculating an Overnight Liquid Culture . After you have bacterial growth, add 500 μL of the overnight culture...bacteria onto an LB agar plate . Grow your bacteria overnight at the appropriate temperature. Growth conditions...page. The next day you will be able to start an overnight culture for plasmid DNA prep the following day...

  14. Plasmid Cloning by PCR

    Type
    Blog Post
    ...amplify your insert DNA. It is important to use a high fidelity polymerase to minimize mutations. The fidelity...

  20. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ... and human embryonic stem cells, they found very high recombination percentages (78-79% after two rounds...

  23. 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 ...

  24. 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...

  30. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ...transfection into mammalian cells is fairly straightforward and the resultant cells can either express ...

  31. 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...

  32. 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...

  35. Antibodies 101: Isotypes

    Type
    Blog Post
    ... an antibody labeled with Fc, Fab, heavy chain, light chain, constant region, and variable region. ...

  38. 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...

  41. 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...

  42. Working with Nuclear Receptors

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

  50. 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...
Showing: 901 - 950 of 978 results