User:Ferdinando Pucci/Notebook/TEMs detection improvement/2008/05/07

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Vector production

Transfer plasmids previously produced:

  • pCCL.SIN-18.Tie2p/e.tTA.PRE
  • pRRL.SIN-18.mTie2e.PPT.mTie2p.GFP.PRE (#416)
  • pRRL.SIN-18.PPT.tetO7.GFP.PRE (#267)

Each vector produced in a 10-cm plate. For Tie2 vectors I used 18ug of transfer plasmid, 15ug for tetO7 (smaller). I used 5ug for both packaging plasmids:

  • If CCL -> pMDLg/p RRE
  • If RRL -> CMV R8.74

Only for CCL systems, added 1mM Na-butyrate in the medium (7ml) 12-14h after transfection.