Registry/Measurement kit/Notebook/2007-7-6

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  • Ligated sequential and double digests of T9002 to 3K3
  • Transforming in MG1655


  • Re-doing BB PCR with vent and phusion
    • Mix for rxn (100uL):
  1. 8uL 2.5mM dNTP
  2. 20uL phusion buffer
  3. 1.5uL f primer
  4. 1.5uL r primer
  5. 1uL DNA
  6. 67uL H2O
  • Just realized that we skipped the ligation step completely (3K3 and 1AK3).
    • Doing that now, discarding PCR product.


  • Re-culturing from plate


  • Checked the sequences; none of them have the promoter.
  • Waiting for primers to insert the promoter.