User:Jeffrey Kim/Notebook/Halogenated natural products from uncultured bacteria/2009/01/23

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TEG JHK TAR

  • Epicentre buffer panel ran against the US and DS targeting sequences for both JHK and TEG pathways as a proof of principle. Good amplifications proudcts found for each amplicon (see panel gel figure). pLLX 8 amplified in bulk with buffer B as optimized earlier (30 cycles, 65C, 0.5 ul 100mM primer).
  • Cloning of pBC 436 is a pain in the @##. Figured out the orientation of the insert was incorrectly annotated. SpeI digest is sufficient. Released the OriT with a PstI digest, and re-cloned it to form the 436 integrative cassette without the oriT.