IGEM:MIT/2005/Experiments: Difference between revisions

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==Experiment Status==
==Experiment Status==


===Jenny===
 
  '''Stock-up MiniPreps'''
  '''Stock-up MiniPreps'''
     [[Image:Pending.gif]] grow up B0015, J13002, E0840
     [[Image:Check.gif]] grow up B0015, J13002, E0840
     [[Image:Pending.gif]] overnight B0015, J13002, E0840
     [[Image:Check.gif]] overnight B0015, J13002, E0840
     [[Image:Pending.gif]] miniprep B0015, J13002, E0840
     [[Image:Check.gif]] miniprep B0015, J13002, E0840
     [[Image:Pending.gif]] glycerol stock J13002
     [[Image:Check.gif]] glycerol stock J13002
 
'''scFv.TT (J07012-14.B0015)'''
    [[Image:Check.gif]] digest B0015 E/X, recipient
    [[Image:Check.gif]] obtain digested scFvs E/S, prefix donor
    [[Image:Check.gif]] gel purify B0015
    [[Image:Check.gif]] ligate J07012-14.B0015
    [[Image:Check.gif]] transform
    [[Image:Check.gif]] overnights for miniprep
    [[Image:Check.gif]] identify correct clones (digest), streak plates
    [[Image:Pending.gif]] overnights for freezer stock
    [[Image:Pending.gif]] freezer stocks scFv.B0015
    [[Image:ScFv for ToxR.jpg|550px]]
 
'''Promoter.RBS.scFv'''
    [[Image:Pending.gif]] PCR start codon to scFv
    [[Image:Pending.gif]] digest Promoter.RBS BB (recipient)
    [[Image:Pending.gif]] obtain digested scFvs (suffix donor)
    [[Image:Pending.gif]] gel purify promoter.RBS
    [[Image:Pending.gif]] ligate promoter.RBS.svFv
    [[Image:Pending.gif]] transform
    [[Image:Pending.gif]] overnights for miniprep
    [[Image:Pending.gif]] identify correct clones (digest), streak plates
    [[Image:Pending.gif]] overnights for freezer stock
    [[Image:Pending.gif]] freezer stocks promoter.RBS.scFv
    [[Image:ScFv for scFvTesting.jpg|550px]]


'''scFvs: 4M5.3, S101A, 4420'''
    [[Image:Check.gif]] biobrick ends
    [[Image:Check.gif]] digest scFv + pSB1AK3-1 plasmid w/ E/S
    [[Image:Check.gif]] gel purify each
    [[Image:Check.gif]] ligate together
    [[Image:Check.gif]] transform
    [[Image:Check.gif]] overnights for miniprep
    [[Image:Check.gif]] identify correct clones (digest)
    [[Image:Check.gif]] overnights for freezer stock
    [[Image:Check.gif]] freezer stocks


  '''FecA (full length)'''
  '''FecA (full length)'''

Latest revision as of 22:34, 14 August 2005

[[../Experimental Flowchart/]]

 indicates done!
 indicates pending...

Experiment Status

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/]]

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