Protocol - pLKO.1 – TRC Cloning Vector
Type
Protocol
...the pattern AA(N 19 ). If no suitable match is found, search for NAR(N 17 )YNN, where N is any nucleotide...and incubate at 37°C, 5% CO 2 for 24 hours. Day 5: n. Harvest media from cells and pool with media from...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...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...