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We narrowed to 992 results for: Ran

Showing: 881 - 900 of 992 results

  8. Colony Formation Titering Assay

    Type
    Protocol
    ...Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots...time as the virus-mediated transduction. For some cell lines, transduction is optimized if the cells are...When the untransduced (negative control) cells have died and colonies are visible in the transduced wells...experiments. Workflow Timeline Day 0: Seed and transduce cells Day 2: Replace media with fresh media containing...to the media, as it can affect viability and transduction efficiency. Titer will vary between cell lines... are seeded the day before transduction. Note that in that case, polybrene should only be added at the...the time of viral transduction, and not during the cell seeding step. Procedure Before beginning a colony...

  9. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... volume of homogenized cell solution is transferred. Transfer cells from the homogenized cell solution...990 µL complete medium. Instead of transferring 0.125 µL, now transfer 100 times that volume, which is 12.5... solution, transfer 12.5 µL of the 1:100 dilution to 10 mL of conditioned medium. Transfer 100 µL of the...generated from A549 cells transduced with lentiCas9-Blast 1 . A549 cells were transduced (MOI = 37) and selected...line from a polyclonal pool of stable cells. Transducing cells with lentivirus results in a heterogenous...heterogeneous cell pool could lead to reduced transgene expression over time, as the lower expressing ...populations that are more likely to retain stable transgene expression. Other methods of generating a monoclonal...

  19. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots...cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment Class...Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other transfection reagents may be used in this protocol, but their...their conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing ...drifted. Filter the solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store... pH to ~8.5 Filter sterilize through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start... Pass and plate the cells the day before the transfection. Thaw a new vial of cells after 30 passages....

  20. Fluorescence Titering Assay

    Type
    Protocol
    ...Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots... were transduced with a range of dilutions of pHAGE-TO-dCas9-3XmCherry . 72 h post transduction, cells...cells transduced F = Fraction of cells with fluorescence D = Dilution Factor V T = Transduction Volume...Workflow Timeline Day 0: Seed 293T cells Day 1: Transduce cells Day 2 (am): Remove media, replace with fresh...to the media, as it can affect viability and transduction efficiency. Titer will vary between cell lines...1468.5 1.5 1:75 20 1478.5 1.5 1:100 15 1483.5 1.5 Untransduced 0 1498.5 1.5 Mix the dilutions well Gently aspirate... one dilution with one well remaining as the untransduced control). Count the cells in the remaining well...
Showing: 881 - 900 of 992 results