IGEM:MIT/2006/Notebook/2006-7-24

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Smell Experiments w/ Isoamyl Acetate

  • Questions that we want to answer:
    1. Does IAA smell like banana? Yes
    2. Does IAA smell strongly in LB?
    3. At what IAA concentration can E. coli "normal" growth?
      • Culture Top10s and add different concentrations of IAA after cells reach OD ~ 0.5

osmY PCR

Done with each pair of new primers meant for PCRing both the short and long osmY promoter region.

  • 45μL PCR SuperMix
  • 2.5μL 0.55ng/μL E. coli genome
  • 1.25μL each respective primer (25 ng/μL)

Did we get transformants using the annealed primers? NO! :(

  • Did you use T4 PNK to 5'-phosphorylate your primers? --Austin 12:20, 24 July 2006 (EDT)

pchB and pchA primers

  • pchB forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA Gat gaa aac tcc cga aga ctg cac - 3'
  • pchB reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT Tat gcg gca ccc cgt gtc tgg cg - 3'
  • pchA forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA Gat gag ccg gct ggc gcc cct gag - 3'
  • pchA reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT Tag gcg acg ccg cgc tgc aag g - 3'
  • pchA mutagenesis forward: 5' - cga ccc att gga ccc gct aca ggt att cgg tgc ctg - 3'
  • pchA mutagenesis reverse: 5' - cag gca ccg aat acc tgt agc ggg tcc aat ggg tcg - 3'
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