... Protocols Lentivirus Production Lentivirus Production You may also like....for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for... Timeline Day 0: Seed 293T packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection...are below passage 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10 6 cells per plate...protocol to generate lentivirus Protocols...Introduction This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T...PES) filter Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid...

... repeat. RRE Rev response element. A.3 Related Products The following plasmids available from Addgene ...control vector containing scrambled shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G ...particles. Note: pLKO.1 can also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG...Incubate at 37°C for 1-2 hours. 4. Run the digestion products on a 1% agarose gel. You should see two fragments...tubes VWR: #87003-290 Note: pLKO.1 could also be packaged using pCMV-dR8.2 dvpr and pCMV-VSVG from the Robert...transfection: 1 μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl... may want to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio...