IGEM:MIT/2005/Wednesday 7/20 Experiment Meeting "Minutes"

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Experimental Planning Meeting:

PURPOSE: Do run through of experimental meeting What do you have What do you need



  1. Permeablizing membrane -- like making cells competent
  2. Attach FFK peptide to antisense agents (RNA) -> lets antisense agent enter cell -> proteins to nucleic acid -> Drew knows things -- proteins that splice out ? -- catch splicing process -- make a tadpole -- protein head and oligo tail
  3. mutant cells: actinomycin (anti biotic) -- strain that takes big things, or just actinomycein?
  4. DNA transport: what exists: conjugation? comEC system (B. subtlis)


  1. Tests this morning didn't go so well
  2. Grew Mc4100 in M9 (doesn't fluoresce), LB
    • no pellet in M9 solution
    • LB cells resuspended in flur. diluted in M9 media
  3. Severe contamination -- Will != sterile
  4. Do the whole set of tests again (can do MC4100 when I do overnights)
  5. Kate says: this expt. was planned at the end of the day, and that was no good at all.
  6. Would like to start dealing with OligoFlurs. --> need assembly



  1. scFv's moving forward



  1. MalE and PhoA -- PhoA had multiple PCR products: TK says raise annealing temperature; Kate: Blast primers against e. coli genome. Though we can just excise band and use that, it makes sense to go forward and do it again


  1. Took protein sequences from cholera toxR -- vibrio species have similar genes, v. fischeri -- also serratia marcescens (don't really believe it is there) -- (sprayed over San Fran)
  2. ATCC -- cholera dna is BL1
    • go ahead and get it -- use this as an exercise
  3. Annotation of translation start on cholera toxR was initially wrong -- there is ambiguity -- ack!! We need to check what we ordered!!



  1. track down fur- strain -- hoping to receive -- how do we get it? RAY = Point of contact
  2. jen and annie talking with samantha to work out recombineering
  3. where do we fuse our scFv? 3 domains: 7 fusions for one domain alone -- working out details -- having a team meeting about this at 4.30 -- should be a team decision -- can we do it in parallel?
  4. FecA in process, FecA prom -- know by end of day tom. if need new primers
  5. FecR and FecI starting pcr



  1. Testing parts, RNA based regulation from IAP 2003



  1. waiting on promoters, going to assemble with gfp when those are ready

Side note: lets talk about risk and danger! Side note: are we overextending ourselves? -gathering materials != doing expt -- might as well gather materials

TO DO: Look into B. Subtleis