IGEM:MIT/2007/Notebook/2007-8-21

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Agenda:

  • Replated Transformations from last week (I14032+B0034+CPX+B0014 in 1AC3, Bl21-e) because old plates potentially compromised from fridge contamination before we could LC them
  • Made our own stock of Cm: NOTE!!! - Endy lab stocks of antibiotics are not for us to use, we need to make our own stock for other antibiotics (Amp, Kan, Tet)
  • More research on Mer Operon - focusing on Tn501: Tried to find exact sequence of pTO40 by backtracking through papers because I wasn't sure how much of merT was included.
  • Basically: They started with Tn501 complete sequence, digested into two fragments by EcoRI cut sites, and ligated into a plasmid called pJOE105 in the place of Tn1702(Which originated from plasmid pJOE100, which came from pBR322). They then cut the new plasmid w/ Tn501, called pJOE114, with EcoRI and AvaI, and ligated that into a commercial pT7-3 plasmid.
  • Since AvaI is located in the first part of the merT gene, only part of merT got transferred into pTO40.
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