... Protocols Lentivirus Production Lentivirus Production You may also like....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...Use cells that are below passage 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10...

...plasmid DNA to mammalian cells Lentivirus Production Produce lentivirus with a polyethyenimine (PEI) transfection...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...lines from a polyclonal pool of stable cells AAV Production in HEK293 Cells Produce adeno-associated virus...

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

... repeat. RRE Rev response element. A.3 Related Products The following plasmids available from Addgene ...Incubate at 37°C for 1-2 hours. 4. Run the digestion products on a 1% agarose gel. You should see two fragments...obtain the ratio that gives you the optimal viral production. d. Create a master mix of FuGENE® 6 transfection...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...