Other ways to cure temperature sensitive ori from E coli?

21 May 2021 0 9K Report

I am attempting to cure pCas plasmid (#62225 in Addgene) (pSC101 ori) in E coli mg1655. I have grown liquid cultures at 42C and plated on selectively. I continued to grow these liquid cultures for >72 hours, periodically plating on selective plates. Still, colonies are growing. I would expect pCas to be cured at 42C in an overnight culture. Any suggestions?

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