Hello everyone,
After lentiviral transduction my I started the antibiotic selection to select the positive clones. The transfected cells did not die suggesting that the transduction worked while the control died after 1 day of puromycin 5ug. However, my plasmid contains the FLAG and when I did the wb I couldn't see the FLAG ( I saw in the positive control so the Ab worked) . Can someone help me? Do you think that transduction works?
I believe in many of these vectors that the antibiotic selection marker and the protein of interest are under separate promoters. It is therefore possible to have antibiotic resistance without the protein of interest expressing. I would sanger sequence your protein of insert to make sure the start codon to stop codon are all correct.
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