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I am trying to clone IRES-puro cassette from pSin-EF2-LIN28-Pur (Addgene) into pLKO.3G (Addgene) replacing EGFP. I got the clone and have transfected cells, but they did not survive puromycin selection even at 0.5 ug/ml concentration. I have no prior experience with IRES. What are the things that need to be looked at while cloning a IRES component? Any help is appreciated. Thanks in advance.
Edited by pratik, 05 March 2010 - 11:44 AM.
