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

Smell Experiments w/ Isoamyl Acetate

 * Questions that we want to answer:
 * Does IAA smell like banana? Yes
 * Does IAA smell strongly in LB?
 * 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.


 * 45&mu;L PCR SuperMix
 * 2.5&mu;L 0.55ng/&mu;L E. coli genome
 * 1.25&mu;L each respective primer (25 ng/&mu;L)

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'