... Protocols Lentivirus Production Lentivirus Production You may also like....This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T cells using...Use cells that are below passage 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10...protocol to generate lentivirus Protocols...lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications...

...Producing Lentiviral Particles E.1 Recommended materials E.2 Protocol for producing lentiviral particles...optimal puromycin concentration F.3 Protocol for lentiviral infection and selection G. Safety H. References...of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the TRC for expression of shRNAs...direct transfection, or can be converted into lentiviral particles for subsequent infection of a target... repeat. RRE Rev response element. A.3 Related Products The following plasmids available from Addgene ...analog). Due to the long terminal repeats found in lentiviral plasmids, Addgene currently recommends using ...Incubate at 37°C for 1-2 hours. 4. Run the digestion products on a 1% agarose gel. You should see two fragments...

...plasmid DNA to mammalian cells Lentivirus Production Produce lentivirus with a polyethyenimine (PEI) transfection...lines from a polyclonal pool of stable cells AAV Production in HEK293 Cells Produce adeno-associated virus...preparation Lentivirus Titration by ddPCR Use ddPCR to measure the physical titer of your lentivirus preparation...transfection protocol Fluorescence Titering Assay for Lentivirus Quantify virus using fluorescence measurements...measurements Colony Formation Titering Assay for Lentivirus Quantify virus by counting antibiotic resistant colonies...colonies Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence into a cell...

...red pH indicator) because the buildup of waste products could be suboptimal for cell growth. This conditioned... 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...