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We narrowed to 988 results for: Ran

Showing: 901 - 920 of 988 results

  8. Western Blot

    Type
    Protocol
    ...Gel Transfer Device. Wet the pre-run gel in deionized water and place it on the transfer membrane of the...protocol uses a dry transfer device but can be adapted for wet and semi-dry transfer approaches. Last Update...well. Assemble the transfer sandwich as follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top... parameters are correct. Transfer for 5–6 min for proteins <30 kDa. Transfer for 8–10 min for proteins...before running your experiment. Remove the membrane from the transfer device and place it in a container with...Block the membrane in blocking buffer for 1 h at RT on a shaking platform. Wash the membrane 3x for 5 ...Incubate the membrane with secondary antibody for 60 min at RT on a shaking platform. Wash the membrane 3x for...

  9. Lentivirus Production

    Type
    Protocol
    ...Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots...lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol. This ...packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection. Remove media, replace...filter Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid... optimized for each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used...drifted. Filter the solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store...

  10. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...drives RNA Polymerase III transcription for generation of shRNA transcripts. cPPT Central polypurine tract...HIV-derivative viruses. For transient knockdown of protein expression, you may transfect plasmid DNA directly...Day 2: b. Perform the transfection in the late afternoon because the transfection mix should only be incubated...it, and does not grant any license, express or implied, under any patent. All transfers of materials from...protocol is provided for your convenience. See “warranty information” in appendix. Table of Contents A....Digesting pLKO.1 TRC-Cloning Vector C.4 Ligating and transforming into bacteria D. Screening for Inserts D.1 Recommended...Sequence of pLKO.1 TRC-Cloning Vector I.2 Recipes I.3 Warranty information Back to Top A. pLKO.1-TRC Cloning ...

  12. A Look at Addgene's QC Process

    Type
    Blog Post
    ...accuracy across all of our plasmids and provide transparency in sequence discrepancies. While Addgene’s QC...

  17. Tips for Technical Support Calls

    Type
    Blog Post
    ...probably difficult for you to ascertain where on this range your question falls. Here are some things to keep...

  18. 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...

  19. 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...
Showing: 901 - 920 of 988 results