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

Experimental Planning Meeting:

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

INPUT
Maxine:
 * 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)

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

HEAD UNIT
Jenny
 * 1) scFv's moving forward

TOXR
Jenny: TK:
 * 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
 * 3) * go ahead and get it -- use this as an exercise
 * 4) Annotation of translation start on cholera toxR was initially wrong -- there is ambiguity -- ack!! We need to check what we ordered!!

FECA
Annie
 * 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

SIGNAL PROCESSING:
Ray:
 * 1) Testing parts, RNA based regulation from IAP 2003

ACTUATOR:
Jen:
 * 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