- Culture cells for protein induction. Just before induction, plate a sample on duplicate plates with and without antibiotic selection. If toxicity is an issue, there will be a significant difference between the number of colonies on the plates. Fewer colonies will be seen on plates containing antibiotic (indicating that the plasmid has been lost) compared to plates without antibiotic.
- Check clone integrity by restriction enzyme analysis and/or sequencing of the ORF.
If toxicity is the problem, add 500 µM L-rhamnose into the selection plates and into the starter culture media.