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  1. Lentivirus Production

    Type
    Protocol
    ... Protocols Lentivirus Production Lentivirus Production You may also ...this protocol to generate lentivirus Protocols...Introduction This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T...lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications...per 10 cm dish* psPAX2 1.3 pmol pMD2.G 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection...PEI is 1:3 (1000 μL total per 10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid...
  2. Lentivirus ddPCR Titration

    Type
    Protocol
    ... of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence... PCR (ddPCR) for lentivirus vector titration. Protocols... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration...Assay AAV Titration by qPCR Addgene Viral Service Lentivirus Guide Virus Blog Posts Introduction This protocol...how to use droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically uses primers...and surfaces with 10% bleach. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please... pipette 10–20 times. Use a new pipette tip to transfer 200 µL of the first dilution into the next dilution...
  3. 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... pool of stable cells. Transducing cells with lentivirus results in a heterogenous polyclonal population...protocol for generating stable cell lines with lentivirus . Day 0: (optional) Seed cells for the generation...protocol for generating stable cell lines with lentivirus . Pro-Tip Because the polyclonal cell pool is...before counting the cells. When you ultimately transfer approximately 50–100 cells to the conditioned ...
  4. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... hours. Day 4: l. Harvest media from cells and transfer to a polypropylene storage tube. The media contains...DS et. al. Cell . 2003. 115:199-208. PubMed . Lentivirus-delivered stable gene silencing by RNAi in primary...1997. 15(9):871-5. PubMed . Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery...license, express or implied, under any patent. All transfers of materials from Addgene to any party are governed...’s Terms of Purchase , and applicable Material Transfer Agreements between the party that deposited the...
Showing: 1 - 4 of 4 results