User:Karmella Haynes/Notebook/ASU courses/2012/01/04

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BME194 Syn Bio Demo Main project page
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1/04/12

  • Choose BioBrick constructs for GFP induction demo
  • Miniprep cultures: GFP1, GFP2, RFP1, RFP2, Banana (5ml each)
  • Plates: Restreak all
  • Plate reader cultures: GFP1, GFP2, RFP1, RFP2, Banana
    • Will do quick and dirty transformations in DH5a-Turbo, and streak plate for Turbo tomorrow

Constructs for the GFP induction experiment
1. GFP1: pLac-GFP + LacI (BBa_K079020), operator LacO2 (K07019)
2. GFP2: pLac-GFP + LacI (BBa_K079021), strong operator LacO1 (K07018)
3. GFP3: pLac-GFP, constitutive, superfolder GFP (from C. Anderson)
4. GFP4: pTet-GFP, constitutive (BBa_I13522) Construct does not express GFP

Cool demo constructs
5. RFP1: pLac-RFP (BBa_J04450)
6. RFP2: pSynth-RFP (BBa_K093012)
7. Banana: pTet-ATF1 (BBa_J45200) (Note: requires isoamyl alcohol to produce smell)

  • All plasmids are Amp resistant
  • Strain: NEB10B (New England Biolabs)

> Start miniprep cultures first, then use 5 ul to inoculate plate reader cultures
> Plate reader cultures (2ml each, overnight):

  1. GFP1
  2. GFP1 + 0.5 mM IPTG
  3. GFP1 + 1.0 mM IPTG
  4. GFP1 + 2.0 mM IPTG
  5. GFP2
  6. GFP2 + 0.5 mM IPTG
  7. GFP2 + 1.0 mM IPTG
  8. GFP2 + 2.0 ug/ml IPTG
  9. GFP4
  10. RFP1
  11. RFP2

Banana test

  1. Banana
  2. Banana + isoamyl alcohol (5ul) Need to order isoamyl alcohol




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