IGEM:MIT/2005/Experiments

[[../Experimental Flowchart/]]

 indicates done!
 indicates pending...

Experiment Status

Promoter.RBS.scFv 
   [[../JennyN Lab Notes#Jenny/]] See Detailed Notes Here.
    PCR start codon to scFv
    digest Promoter.RBS BB (recipient)
    digest scFvs (suffix donor)
    gel purify 
    ligate promoter.RBS.svFv
    transform
    overnights for miniprep
    identify correct clones (digest), streak plates
    overnights for freezer stock
    freezer stocks promoter.RBS.scFv
   

Stock-up MiniPreps
    grow up B0015, J13002, E0840
    overnight B0015, J13002, E0840
    miniprep B0015, J13002, E0840
    glycerol stock J13002

FecA (full length)
    PCR bb ends
    digest FecA + pSB1AK3-1 plasmid w/ E/S
    gel purify each
    ligate together
    transform
    overnights for miniprep
    identify correct clones
    overnights for freezer stock
    freezer stocks

Jessica

MalE
    PCR
    digest MalE + pSB1AK3-1 plasmid w/ E/S 
    gel purify each
    ligate together
    transform
    identify correct clones

PhoA
    PCR with higher annealing temperature
    digest PhoA+ pSB1AK3-1 plasmid w/ E/S 
    gel purify each
    ligate together
    transform
    identify correct clones

PhoA'
    QuikChange

Annie

FecA promoter
    PCR with current primers with higher annealing temperature
    PCR with newly designed primers
    digest FecApro + pSB1AK3-1 plasmid w/ E/S 
    gel purify each
    ligate together
    transform
    identify correct clones (digest)
    freezer stock

FecI
    PCR bb end
    digest FecI + pSB1AK3-1 plasmid w/ E/S
    gel purify each
    ligate together
    transform
    identify correct clones (digest)
    freezer stock

FecR
    PCR bb end
    digest FecR + pSB1AK3-1 plasmid w/ E/S
    gel purify each
    ligate together
    transform
    identify correct clones (digest)
    freezer stock

FecA'
    QuikChange

FecR'
    QuikChange

Project Experiments

  Input Device: [[../Input: Ligand#Experiments/|fluorescein]]
  Head Receiver Unit: [[../Receiver Head Unit: scFv#Experiments/|anti-fluorescein scFv]]
  Receiver 1: [[../Receiver 1: ToxR#Experiments/|ToxR]]
  Receiver 2: [[../Sensor 2#Experiments/|FecA]] 
  Signal Processor: [[../Signal processing#Experiments/|Exp. Here]]
  Actuator: [[../Actuator#Experiments/|Exp. Here]]

[[../Experimental Timeline/]]